ࡱ> |yz{ [bjbj >>[rrBBBBBVVVV,V{]*&4 4 4 4 #!#!#!\\\\\\\,_Wb\\-B#!#!#!#!#!\$BB4 4 ]6$$$#!&B4 B4 \$#!\$$2ILPP c湀I# N.\?]<{]NNb$b\PP$PPB^U<#!#!#!\\$#!#!#!{]b#!#!#!#!#!#!#!#!#!r :   True rifampicin resistance missed by the MGIT: prevalence of this pheno/genotype in the UK and Ireland after 18 month surveillance Ximena Gonzalo1, 2, Pauline Claxton3, Tim Brown1, Lorraine Montgomery4, Margaret Fitzgibbon4, Ian Laurenson3, Francis Drobniewski1, 2 National Mycobacterium Reference Laboratory, Public Health England, 2 Newark Street, London E1 2AT, UK Department of Infectious Diseases, Imperial College, The Commonwealth Building, The Hammersmith Hospital, Du Cane Road, London W12 0NN, UK Scottish Mycobacteria Reference Laboratory, Clinical Microbiology, Royal Infirmary of Edinburgh, 51 Little France Crescent, Edinburgh, EH16 4SA The Irish Mycobacteria Reference Laboratory, St. James's Hospital, James's Street, Dublin 8, Ireland Corresponding author: Francis Drobniewski,  HYPERLINK "mailto:f.drobniewski@imperial.ac.uk" f.drobniewski@imperial.ac.uk, Department of Infectious Diseases, Imperial College, The Commonwealth Building, The Hammersmith Hospital, Du Cane Road, London W12 0NN, UK. Phone number: +44 020 7589 5111 Intended article format: Original article Running title: Rifampicin resistance missed by the MGIT Abstract Objectives: To characterise rifampicin resistant strains missed by the MGIT 960 system but not by egg-based media in the UK and Ireland and to ascertain their prevalence. Methods: All strains sent for second line susceptibility testing were prospectively collected. Drug Susceptibility Testing was performed by Resistance Ratio (RR), Proportion Method (PM), MGIT 960 and MIC determination by microdilution. Rifampicin resistance conferring mutations were detected with line probe assays and sequencing. At the end of the study period, retrospective archived strains from 2010 to 2014 showing key mutations were analysed phenotypically (by MGIT 960 and RR) and genotypically. Results: 17/7234 prospective isolates were included. All of them were susceptible by MGIT. One was borderline by RR (MIC to RIF of 4mg/L) and was resistant by PM. 8/17 were resistant and 8/17 were highly resistant on RR. These 16 isolates had MICs between 1 and 8 mg/L on microdilution. With PM, 16/17 were susceptible to rifampicin. 17/17 had mutations in the rpoB gene. D516Y was the mutation most frequently found (13/17). Retrospectively 10 additional strains with key genotypes were found in our collection: 6/10 were susceptible in the MGIT and resistant in RR. Of the 27 studied strains, the MGIT only detected resistance in 4. Conclusions: Rifampicin resistance is missed by the MGIT system. In the UK and Ireland the prevalence of these strains is low. The introduction of routine molecular testing would detect false susceptibility. Further research is needed to ascertain the role of these strains in clinical failure and their prevalence in other settings. Introduction Rifampicin is the backbone of anti-tuberculosis treatment and resistance in Mycobacterium tuberculosis is associated with increased mortality.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.5539/gjhs.v5n6p60", "ISSN" : "1916-9736", "PMID" : "24171875", "abstract" : "BACKGROUND: Drug resistance substantially increases tuberculosis (TB) mortality. This study aimed to describe the prevalence of mycobacterial drug resistance pattern and association of common resistance patterns with TB mortality in Thailand.\n\nMETHOD: A retrospective cohort study was conducted using TB surveillance data. A total of 9,518 culture-confirmed, pulmonary TB patients registered from 1 October 2004 to 31 December 2008 from the Thailand TB Active Surveillance Network were included in this study. Patients were followed up until TB treatment completion or death. Mycobacterial drug resistance patterns were categorized as pan-susceptible, rifampicin resistance, isoniazid monoresistance, and ethambutol/streptomycin resistance. Drug susceptibility testing (DST) was determined by Mycobacterial Growth Indicator Tube (MGIT) liquid culture systems. Survival analysis was applied.\n\nRESULT: Isoniazid monoresistance was the most common pattern, while rifampicin resistance had the largest impact on mortality. Cox regression analysis showed a significantly higher risk of death among patients with rifampicin resistance (adjusted hazard ratio (aHR) 1.9, 95% confident interval (CI), 1.5-2.5) and isoniazid monoresistance (aHR 1.4, 95% CI 1.1-1.7) than those with pan-susceptible group after adjustment for age, nationality, human immunodeficiency virus (HIV) and antiretroviral therapy (ART) status, diabetes mellitus, cavitary disease on chest x-ray, treatment observation, and province. HIV co-infection was associated with higher mortality in patients both on ART (aHR 1.9, 95% CI 1.5-2.5) and not on ART (aHR 8.1, 95% CI 6.8-9.8).\n\nCONCLUSION: Rifampicin resistance and isoniazid monoresistance were associated with increased TB mortality. HIV-coinfection was associated with a higher risk of death including among those taking antiretroviral therapy.", "author" : [ { "dropping-particle" : "", "family" : "Anuwatnonthakate", "given" : "Amornrat", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Whitehead", "given" : "Sara J", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Varma", "given" : "Jay K", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Silachamroon", "given" : "Udomsak", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Kasetjaroen", "given" : "Yuthichai", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Moolphate", "given" : "Saiyud", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Limsomboon", "given" : "Pranom", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Inyaphong", "given" : "Jiraphun", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Suriyon", "given" : "Narin", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Kavinum", "given" : "Suporn", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Chiengson", "given" : "Navarat", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Tunteerapat", "given" : "Phatchara", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Kaewkungwal", "given" : "Jaranit", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Global journal of health science", "id" : "ITEM-1", "issue" : "6", "issued" : { "date-parts" : [ [ "2013", "11" ] ] }, "page" : "60-72", "title" : "Effect of mycobacterial drug resistance patterns on patients' survival: a cohort study in Thailand.", "type" : "article-journal", "volume" : "5" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=71126211-343f-4c60-b295-6e9456331820" ] } ], "mendeley" : { "formattedCitation" : "[1]", "plainTextFormattedCitation" : "[1]", "previouslyFormattedCitation" : "[1]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[1] Accurately determining susceptibility is of paramount importance, not only to establish the treatment regimen and duration but also to improve cure rates and survival.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "ISSN" : "1399-3003", "author" : [ { "dropping-particle" : "", "family" : "World Health Organization", "given" : "", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "2011", "9" ] ] }, "title" : "WHO guidelines for the programmatic management of drug-resistant tuberculosis: 2011 update.", "type" : "article-journal" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=02f8f7cb-248c-4ce9-89ac-9293c50ce48c" ] }, { "id" : "ITEM-2", "itemData" : { "author" : [ { "dropping-particle" : "", "family" : "World Health Organization", "given" : "", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "4th Edition", "id" : "ITEM-2", "issued" : { "date-parts" : [ [ "2009" ] ] }, "title" : "Treatment of tuberculosis. Guidelines.", "type" : "paper-conference" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=a6178f4c-04e9-44f9-82bd-9cd131810c99" ] } ], "mendeley" : { "formattedCitation" : "[2,3]", "plainTextFormattedCitation" : "[2,3]", "previouslyFormattedCitation" : "[2,3]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[2,3] Currently, a range of genotypic and phenotypic susceptibility methods exist to detect rifampicin resistance. Even though molecular detection of rifampicin resistance is recommended, not all laboratories in the UK and Ireland use them on all M. tuberculosis cases or have them amongst their locally available methods. In the United Kingdom National Mycobacterium Reference Laboratory (NMRL), molecular detection of rifampicin resistance is available but it is not performed universally; it is done on request or upon detection of either risk factors for drug resistance or phenotypic resistance. At the Scottish Mycobacteria Reference Laboratory (SMRL) direct molecular testing for rifampicin and usually isoniazid is carried out on specimens which are M. tuberculosis complex qPCR first film positive respiratory specimens and on request.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1099/jmm.0.043166-0", "ISSN" : "1473-5644", "PMID" : "22790204", "abstract" : "An in-house IS6110 real-time PCR (IH IS6110), MTB Q-PCR Alert (Q-PCR) and GenoType MTBDRplus (MTBDR; Hain Lifescience) were compared for the direct detection of Mycobacterium tuberculosis complex (MTBC) in 87 specimens following automated NucliSENS easyMAG DNA extraction. This included 82 first smear-positive specimens and three smear-negative specimens. Another in-house real-time PCR with a Mycobacterium genus-specific probe for the internal transcribed spacer (ITS) region (IH ITS) was used to allow a full comparison with culture results. The sensitivities of IH IS6110, Q-PCR, MTBDR and IH ITS for MTBC detection were 100, 92, 87 and 87\u200a%, respectively, compared with culture. Both IS6110-based real-time PCRs (in-house and Q-PCR) were similar in performance, with 91.2\u200a% concordant results for MTBC detection. Inhibition rates were low, with zero to three specimens producing uninterpretable results. However, the Q-PCR failed to detect MTBC in five samples that were smear negative or had few acid-fast bacilli (one to 10 bacilli in 10 microscopic fields) detected by IH IS6110. IH ITS was the least sensitive assay but may be useful when used in conjunction with IS6110 PCR results to determine the presence of non-tuberculous mycobacteria in smear-negative specimens. None of the real-time PCR assays tested provided drug-resistance data. It was concluded that an IH IS6110 assay could easily be incorporated into the workflow of a diagnostic laboratory for rapid and accurate identification of MTBC from clinical specimens. The inclusion of an internal control and amplification of an ITS target enhance the diagnostic utility of the test.", "author" : [ { "dropping-particle" : "", "family" : "Seagar", "given" : "Amie-Louise", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Neish", "given" : "Barry", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Laurenson", "given" : "Ian F", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of medical microbiology", "id" : "ITEM-1", "issue" : "Pt 10", "issued" : { "date-parts" : [ [ "2012", "10" ] ] }, "page" : "1459-64", "title" : "Comparison of two in-house real-time PCR assays with MTB Q-PCR Alert and GenoType MTBDRplus for the rapid detection of mycobacteria in clinical specimens.", "type" : "article-journal", "volume" : "61" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=612af408-eb2d-4a31-a419-6ad84234ec53" ] } ], "mendeley" : { "formattedCitation" : "[4]", "plainTextFormattedCitation" : "[4]", "previouslyFormattedCitation" : "[4]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[4] Similarly at the Irish Mycobacteria Reference Laboratory (IMRL), direct molecular testing for rifampicin resistance is performed on request and on all new smear positive respiratory specimens. Culture using the automated Mycobacteria Growth Indicator Tube (MGIT) is one of the gold standard methodologies for phenotypic susceptibility testing endorsed by the World Health Organisation.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "author" : [ { "dropping-particle" : "", "family" : "World Health Organization", "given" : "", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "0" ] ] }, "title" : "Guidelines for surveillance of drug resistance in tuberculosis", "type" : "article-journal" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=56b348fc-95cf-424d-9689-99bb3cf014f3" ] } ], "mendeley" : { "formattedCitation" : "[5]", "plainTextFormattedCitation" : "[5]", "previouslyFormattedCitation" : "[5]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[5] Its introduction improved turn-around times compared to solid media and it has been adopted by tuberculosis laboratories worldwide. However, reports that the MGIT system misses some strains identified as resistant to rifampicin by other culture based or molecular methods have started to emerge.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1128/JCM.02741-12", "ISSN" : "1098-660X", "PMID" : "23761146", "abstract" : "WHO-endorsed phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are assumed to be the gold standard for identifying rifampin (RMP) resistance. However, previous results indicated that low-level, yet probably clinically relevant, RMP resistance linked to specific rpoB mutations is easily missed by some growth-based methods. We aimed to compare the level of resistance detected on L\u00f6wenstein-Jensen (LJ) medium with resistance detected by the Bactec MGIT 960 automated DST (MGIT-DST) system for various rpoB mutants. Full agreement between LJ and MGIT-DST was observed for mutations located at codons 513 (Lys or Pro) and 531 (Leu, Trp), which were always resistant by both methods. For mutations 511Pro, 516Tyr, 533Pro, 572Phe, and several 526 mutations, LJ and MGIT results were highly discordant, with MGIT-DST failing to give a result or declaring the strains susceptible. Our data show that phenotypic RMP resistance testing of M. tuberculosis is not a binary phenomenon for some rpoB mutations and that the widely used automated MGIT 960 system is prone to miss some RMP resistance-conferring mutations, while careful DST on LJ missed hardly any. Given the association of these mutations with poor clinical outcome, our findings suggest that the gold standard for rifampin resistance should be reconsidered, in order to address the present confusion caused by discrepancies between phenotypic and genotypic results. The impacts of these mutations will depend on the frequency of their occurrence, which may vary from one setting to another.", "author" : [ { "dropping-particle" : "", "family" : "Rigouts", "given" : "Leen", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Gumusboga", "given" : "Mourad", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rijk", "given" : "Willem Bram", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Nduwamahoro", "given" : "Elie", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Uwizeye", "given" : "C\u00e9cile", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jong", "given" : "Bouke", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Deun", "given" : "Armand", "non-dropping-particle" : "Van", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-1", "issue" : "8", "issued" : { "date-parts" : [ [ "2013", "8" ] ] }, "page" : "2641-5", "title" : "Rifampin resistance missed in automated liquid culture system for Mycobacterium tuberculosis isolates with specific rpoB mutations.", "type" : "article-journal", "volume" : "51" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=6e35a569-8c72-417e-ac7a-a65b1473fefd" ] } ], "mendeley" : { "formattedCitation" : "[6]", "plainTextFormattedCitation" : "[6]", "previouslyFormattedCitation" : "[6]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[6] Not only was true resistance missed by the automated method but these errors did not seem to occur with the proportion method in Lowenstein-Jensen (LJ) medium. At the National Mycobacterium Reference Laboratory, the MGIT was used only for isolation, and for second line drug testing (amikacin, capreomycin, kanamycin, ethionamide, prothionamide, linezolid, moxifloxacin and ofloxacin), with first line drugs and PAS tested using the resistance ratio method (RR) on LJ media exclusively. However, other reference centres in the UK and Ireland perform first line susceptibility testing using MGIT. Therefore, true rifampicin resistance might be missed. Surveillance was started for a period of 18 months in both the United Kingdom and the Republic of Ireland to ascertain the prevalence of this phenomenon in the circulating strains. We report the result of that surveillance. Methods All samples received by the study reference centres for identification and susceptibility, received during the 18 months July 2012 December 2013 inclusive and identified as M. tuberculosis were included. All of those requiring second line drug susceptibility testing (DST) either because of resistance to a first line drug or a clinicians request or detected as rifampicin resistant by direct molecular testing were further analysed by all the methods described below. For the second line drugs, two MGIT tubes containing isoniazid and rifampicin were added. Samples came from the NMRL, the SMRL the IMRL. Susceptibility testing was performed as follows: Resistance ratio (RR): briefly, for every new batch of in-house prepared LwensteinJensen medium, 3 known strains plus H37Rv were used to determine the modal susceptibility pattern against which the clinical strains were tested. Three different concentrations of rifampicin were tested (5, 10 and 20 mg/L) and results, read at 14, 21 and 28 days, were reported as susceptible, borderline, resistant or highly resistant after the following calculation: RR= Test MIC/Modal control MIC QUOTE  ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "ISBN" : "0781749832", "abstract" : "The Fifth Edition of this highly acclaimed reference describes state-of-the-art laboratory methods for evaluating the efficacy of antibiotics and the mechanisms of bacterial resistance. Coverage includes new molecular techniques for antibiotic testing, the new discipline of pharmacogenomics, and use of computers to monitor resistance trends. New chapters focus on the breakpoint, molecular techniques for susceptibility testing, susceptibility testing of HIV and hepatitis agents to antiviral drugs, and disinfectant and antiseptic testing. Sixteen new antimicrobial agents most FDA-approved, others German and UK-approved are examined. More than 200 tables provide information on every organism, every antimicrobial agent, and drug concentrations in blood, organs, and tissues.", "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "2005" ] ] }, "number-of-pages" : "889", "publisher" : "Lippincott Williams & Wilkins", "title" : "Antibiotics in Laboratory Medicine", "type" : "book" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=aa05d9b1-d560-4738-9c69-e6570c06aea6" ] } ], "mendeley" : { "formattedCitation" : "[7]", "plainTextFormattedCitation" : "[7]", "previouslyFormattedCitation" : "[7]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[7] Proportion method on Middlebrook 7H10 (PM): critical concentration of rifampicin 1 mg/L. Critical proportion 1%. Plates were read at 14, 21 and 28 days. The valid read was the one obtained when the growth control plate showed more than 50 colonies. Results are either susceptible or resistant.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "ISSN" : "0042-9686", "PMID" : "5309084", "abstract" : "In a paper arising out of an informal international consultation of specialists in the bacteriology of tuberculosis held in 1961, an attempt was made to formulate criteria, and specify technical procedures, for reliable tests of sensitivity (the absolute-concentration method, the resistance-ratio method and the proportion method) to the 3 main antituberculosis drugs (isoniazid, streptomycin and p-aminosalicylic acid). Seven years later, a further consultation was held to review the latest developments in the field and to suggest how sensitivity tests might be put to practical use in tuberculosis control programmes. The participants reached agreement on how to define drug sensitivity and resistance, and stressed the importance of using a discrimination approach to the calibration of sensitivity tests. Their views are contained in the present paper, which also includes descriptions of the sensitivity tests used by the Medical Research Council of Great Britain for first- and second-line drugs (minimal inhibitory concentration and resistance-ratio methods), the two main variants of the proportion method developed by the Institut Pasteur, Paris, and a method for calibrating sensitivity tests.", "author" : [ { "dropping-particle" : "", "family" : "Canetti", "given" : "G", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Fox", "given" : "W", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Khomenko", "given" : "A", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Mahler", "given" : "H T", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Menon", "given" : "N K", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Mitchison", "given" : "D A", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rist", "given" : "N", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Smelev", "given" : "N A", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Bulletin of the World Health Organization", "id" : "ITEM-1", "issue" : "1", "issued" : { "date-parts" : [ [ "1969", "1" ] ] }, "page" : "21-43", "title" : "Advances in techniques of testing mycobacterial drug sensitivity, and the use of sensitivity tests in tuberculosis control programmes.", "type" : "article-journal", "volume" : "41" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=0de5a0a1-a1f6-42f8-a3fc-8e9f2165d47c" ] } ], "mendeley" : { "formattedCitation" : "[8]", "plainTextFormattedCitation" : "[8]", "previouslyFormattedCitation" : "[8]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[8] MGIT 960(Becton Dickinson Microbiology Systems, Sparks, Md.) susceptibility testing according to manufacturers instructions. Results are either susceptible or resistant.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "author" : [ { "dropping-particle" : "", "family" : "Palomino", "given" : "J. C.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Traore", "given" : "H.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Fissette", "given" : "K.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Portaels", "given" : "F.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "0" ] ] }, "publisher" : "International Union Against Tuberculosis and Lung Disease", "title" : "Evaluation of Mycobacteria Growth Indicator Tube (MGIT) for drug susceptibility testing of Mycobacterium tuberculosis", "type" : "article-journal" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=f1c231a9-dd2c-4a54-b1c9-a0d44037dc51" ] } ], "mendeley" : { "formattedCitation" : "[9]", "plainTextFormattedCitation" : "[9]", "previouslyFormattedCitation" : "[9]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[9] MIC determination by microdilution (TREK Diagnostic Systems, Cleveland, OH) Concentrations tested were 0.12-16 mg/L. Breakpoint is 1mg/L (d"1mg/L is susceptible).ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1007/s10096-011-1382-z", "ISSN" : "1435-4373", "PMID" : "21866324", "abstract" : "The agar proportion method (APM) for determining Mycobacterium tuberculosis susceptibilities is a qualitative method that requires 21 days in order to produce the results. The Sensititre method allows for a quantitative assessment. Our objective was to compare the accuracy, time to results, and ease of use of the Sensititre method to the APM. 7H10 plates in the APM and 96-well microtiter dry MYCOTB panels containing 12 antibiotics at full dilution ranges in the Sensititre method were inoculated with M. tuberculosis and read for colony growth. Thirty-seven clinical isolates were tested using both methods and 26 challenge strains of blinded susceptibilities were tested using the Sensititre method only. The Sensititre method displayed 99.3% concordance with the APM. The APM provided reliable results on day 21, whereas the Sensititre method displayed consistent results by day 10. The Sensititre method provides a more rapid, quantitative, and efficient method of testing both first- and second-line drugs when compared to the gold standard. It will give clinicians a sense of the degree of susceptibility, thus, guiding the therapeutic decision-making process. Furthermore, the microwell plate format without the need for instrumentation will allow its use in resource-poor settings.", "author" : [ { "dropping-particle" : "", "family" : "Abuali", "given" : "M M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Katariwala", "given" : "R", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "LaBombardi", "given" : "V J", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology", "id" : "ITEM-1", "issue" : "5", "issued" : { "date-parts" : [ [ "2012", "5" ] ] }, "page" : "835-9", "title" : "A comparison of the Sensititre\u00ae MYCOTB panel and the agar proportion method for the susceptibility testing of Mycobacterium tuberculosis.", "type" : "article-journal", "volume" : "31" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=fec311e6-682e-49e7-a0ba-1e7ca76bf561" ] }, { "id" : "ITEM-2", "itemData" : { "DOI" : "10.1210/jc.2010-0195", "ISSN" : "1945-7197", "PMID" : "20427486", "abstract" : "BACKGROUND: Serum 25-hydroxyvitamin D (25OHD) is a key factor in determining monocyte induction of the antimicrobial protein cathelicidin, which requires intracrine conversion of 25OHD to 1,25-dihydroxyvitamin D [1,25(OH)(2)D]. Both vitamin D metabolites circulate bound to vitamin D-binding protein (DBP), but the effect of this on induction of monocyte cathelicidin remains unclear.\n\nMETHODS: Human monocytes were cultured in medium containing 1) serum from DBP knockout (DBP(-/-)) or DBP(+/-) mice, 2) serum-free defined supplement reconstituted with DBP or albumin (control), and 3) human serum with different DBP [group-specific component [Gc]] genotypes with varying affinities for vitamin D metabolites. In each case, response to added 1,25(OH)(2)D(3) or 25OHD(3) was determined by measuring expression of mRNA for cathelicidin and 24-hydroxylase. Monocyte internalization of DBP was assessed by fluorescent tagging followed by microscopic and flow cytometric analysis of tagged DBP.\n\nRESULTS: Monocytes cultured in DBP(-/-) serum showed more potent induction of cathelicidin by 25OHD(3) or 1,25(OH)(2)D(3) when compared with DBP(+/-) serum. Likewise, DBP added to serum-free medium attenuated 25OHD(3)/1,25(OH)(2)D(3) responses. Fluorescently tagged DBP showed low-level uptake by monocytes, but this did not appear to involve a megalin-mediated mechanism. Human serum containing low-affinity Gc2-1S or Gc2-2, respectively, supported 2.75-fold (P = 0.003) and 2.43-fold (P = 0.016) higher induction of cathelicidin by 25OHD relative to cells cultured with high affinity Gc1F-1F.\n\nCONCLUSION: These data indicate that DBP plays a pivotal role in regulating the bioavailablity of 25OHD to monocytes. Vitamin D-dependent antimicrobial responses are therefore likely to be strongly influenced by DBP polymorphisms.", "author" : [ { "dropping-particle" : "", "family" : "Chun", "given" : "Rene F", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Lauridsen", "given" : "Anna L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Suon", "given" : "Lizabeth", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Zella", "given" : "Lee a", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Pike", "given" : "J Wesley", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Modlin", "given" : "Robert L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Martineau", "given" : "Adrian R", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Wilkinson", "given" : "Robert J", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Adams", "given" : "John", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Hewison", "given" : "Martin", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "The Journal of clinical endocrinology and metabolism", "id" : "ITEM-2", "issue" : "7", "issued" : { "date-parts" : [ [ "2010", "7" ] ] }, "page" : "3368-76", "title" : "Vitamin D-binding protein directs monocyte responses to 25-hydroxy- and 1,25-dihydroxyvitamin D.", "type" : "article-journal", "volume" : "95" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=35d515bd-af14-42d3-bd9e-36b9133f2cf2" ] } ], "mendeley" : { "formattedCitation" : "[10,11]", "plainTextFormattedCitation" : "[10,11]", "previouslyFormattedCitation" : "[10,11]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[10,11] Variable Number Tandem Repeat (VNTR) was performed in all strains as part of routine analysis, as per Brown et al.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1007/978-1-59745-207-6_25", "ISSN" : "1940-6029", "PMID" : "20560063", "abstract" : "DNA-based typing has contributed to the understanding of M. tuberculosis epidemiology and evolution. IS6110 RFLP was the first method described and has been used in many epidemiologic investigations. Technological difficulties have hampered the widespread establishment of this method, and it has been found to be of little use in evolutionary studies. PCR-based methods such as spoligotyping and variable number tandem repeat (VNTR) analysis largely overcome these difficulties. Spoligotyping alone is of limited value in epidemiologic investigations due to low discrimination but can be useful in evolutionary studies. Panels of VNTR loci selected from the 59 polymorphic VNTRs described to date have been shown to be useful in both epidemiologic and evolutionary studies. A VNTR type is identified by, first, amplifying a series of PCR fragments each encompassing a different VNTR locus and, second, determining the PCR fragment sizes from which the number of repeats present is calculated. The repeat number present at a series of loci is used as numerical code to describe a type. This chapter describes a high-throughput automated method for VNTR analysis at 15 loci using a capillary fragment analyzer and a manual method using agarose gel analysis.", "author" : [ { "dropping-particle" : "", "family" : "Brown", "given" : "T J", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Nikolayevskyy", "given" : "V N", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Drobniewski", "given" : "F A", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Methods in molecular biology (Clifton, N.J.)", "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "2009", "1" ] ] }, "page" : "371-94", "title" : "Typing Mycobacterium tuberculosis using variable number tandem repeat analysis.", "type" : "article-journal", "volume" : "465" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=ffd099fb-f74f-43f6-87aa-41a55914d357" ] } ], "mendeley" : { "formattedCitation" : "[12]", "plainTextFormattedCitation" : "[12]", "previouslyFormattedCitation" : "[12]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[12] Mutations conferring resistance to rifampicin were detected using line probe assays (HAIN GenoType MTB DR plus (Hain Lifescience GmbH, Nehren, Germany) and by rpoB sequencing using the CEQ 8000 Genetic Analysis System (Beckman Coulter, Inc., Fullerton, CA, USA) from nucleotides 507 to 533. Performance of the genotypic analysis was blinded to result of the phenotypic MGIT analysis. Additionally, at the end of the study period (from 01/07/2012 until 31/12/2013) a retrospective interrogation of the NMRL mutation database was conducted of all the strains from 2010 until 2014. Strains showing the mutations observed in the prospectively collected study (D516Y, H525Y, L511P+M515I) were subcultured from the archive and analysed phenotypically (RR and MGIT) and again genotypically as stated above. Both in vitro and genetic susceptibility testing are part of the routine work up of samples and the VNTR analysis is part of the public health and epidemiology commintment of the reference services provided by the centres. All the results were made available on real time to clinicians and the public health authority. No clinical information was available or used. No ethical approval was required. Results From the 18 month period July 2012- December 2013 inclusive 7234 isolates were identified as M. tuberculosis and had first line DST performed in the UK and Ireland. 346 M. tuberculosis cultures had first line susceptibility performed at the IMRL (9 sent to NMRL for second line drug testing), 422 at the SMRL (3 were sent to NMRL for further testing) and 3959 positive TB cultures had first line drug susceptibility testing performed at the NMRL. Additionally, 38 strains whose first line DSTs were performed in other PHE regional centres (total number processed 2557), were sent to the NMRL for second line testing; 323 isolates were re-tested as per protocol. After repeat testing 17 isolates from 13 patients were borderline, resistant or highly resistant to rifampicin on RR testing and were susceptible on the MGIT system. Six came from London, 3 from the West Midlands, 2 from Scotland and 2 from Ireland. For a full result panel, see online supplement Table 1. Eight isolates were resistant and 8 were highly resistant on RR. 1 of 17 isolates was borderline resistant by RR. This isolate had an MIC to RIF of 4 mg/L(R) and was also resistant by PM on 7H10. Of the resistant strains, 5 had an MIC of 1 mg/L (S), one of 2 mg/L (R) and 2 of 4 mg/L (R). Of the highly resistant strains, 2 showed an MIC of 1 mg/L (S), 2 of 2 mg/L (R), 2 of 4 mg/L (R) and 2 of 8 mg/L (R). Interestingly, as stated in the previous paragraph, 7 isolates were classed as susceptible (MIC 1 mg/L) following current recommendations where the resistant MIC is >1mg/L.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1128/JCM.02048-12", "ISSN" : "1098-660X", "PMID" : "22895034", "abstract" : "The Sensititre MycoTB plate (TREK Diagnostic Systems, Cleveland, OH) uses a microtiter plate MIC format for susceptibility testing of Mycobacterium tuberculosis complex isolates against first- and second-line antituberculosis agents. Categorical agreement versus the agar proportion method for 122 M. tuberculosis complex isolates was 94% to 100%.", "author" : [ { "dropping-particle" : "", "family" : "Hall", "given" : "Leslie", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jude", "given" : "Kurt P", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Clark", "given" : "Shirley L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Dionne", "given" : "Kim", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Merson", "given" : "Ryan", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Boyer", "given" : "Ana", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Parrish", "given" : "Nicole M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Wengenack", "given" : "Nancy L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-1", "issue" : "11", "issued" : { "date-parts" : [ [ "2012", "11" ] ] }, "page" : "3732-4", "title" : "Evaluation of the Sensititre MycoTB plate for susceptibility testing of the Mycobacterium tuberculosis complex against first- and second-line agents.", "type" : "article-journal", "volume" : "50" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=968c8c2a-2773-491e-a29a-fa678555299f" ] } ], "mendeley" : { "formattedCitation" : "[13]", "plainTextFormattedCitation" : "[13]", "previouslyFormattedCitation" : "[13]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[13] On PM using 7H10 media, 8 were susceptible at week 2 while 9 could not be read since the GC had less than 50 colonies. At week 3, 5 additional isolates showed susceptibility and 3 were resistant while only one remained not ready to be read due to GC poor growth. On week 4, the last isolate was ready and was resistant. So, after the first valid reading as per protocol 16 out of 17 isolates were susceptible on 7H10 media. On re-incubation off protocol (after the valid read has been obtained), 7 isolates that were previously defined as susceptible became resistant by week four of incubation. One isolate was contaminated on re-incubation. All 17 isolates had mutations in the rpoB gene, detected by the Hain line probe assay, by absence of the wild type bands and/or the presence of mutant bands. Ten isolates were multidrug resistant while 7 were resistant to rifampicin alone D516Y was the mutation most frequently found. It was present in 13 isolates. H526L was found only once and the combination of L511Pplus M515I was present in 3 isolates. Two isolates belonged to the Beijing lineage whereas 5 were Euro American and 10 were CAS. There were 3 pairs belonging to the same patients, but otherwise no clusters defined by VNTR. When looking for the genotype containing the stated mutations, 10 new isolates were found in our collection: 4 isolates containing the D516Y mutation were resistant to RIF in the MGIT. However, six of them behaved similarly, being susceptible in the MGIT and R or HR by RR (online supplement Table 2). No MICs or proportion method in 7H10 were available for these isolates since they were analysed retrospectively and were not part of the prospective study. Of the 27 isolates containing the above mutations, only 4 were identified as resistant by the MGIT system. Discussion Previous reports that a proportion of true rifampicin resistance was missed by the MGIT system were confirmed by this study.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1128/JCM.02741-12", "ISSN" : "1098-660X", "PMID" : "23761146", "abstract" : "WHO-endorsed phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are assumed to be the gold standard for identifying rifampin (RMP) resistance. However, previous results indicated that low-level, yet probably clinically relevant, RMP resistance linked to specific rpoB mutations is easily missed by some growth-based methods. We aimed to compare the level of resistance detected on L\u00f6wenstein-Jensen (LJ) medium with resistance detected by the Bactec MGIT 960 automated DST (MGIT-DST) system for various rpoB mutants. Full agreement between LJ and MGIT-DST was observed for mutations located at codons 513 (Lys or Pro) and 531 (Leu, Trp), which were always resistant by both methods. For mutations 511Pro, 516Tyr, 533Pro, 572Phe, and several 526 mutations, LJ and MGIT results were highly discordant, with MGIT-DST failing to give a result or declaring the strains susceptible. Our data show that phenotypic RMP resistance testing of M. tuberculosis is not a binary phenomenon for some rpoB mutations and that the widely used automated MGIT 960 system is prone to miss some RMP resistance-conferring mutations, while careful DST on LJ missed hardly any. Given the association of these mutations with poor clinical outcome, our findings suggest that the gold standard for rifampin resistance should be reconsidered, in order to address the present confusion caused by discrepancies between phenotypic and genotypic results. The impacts of these mutations will depend on the frequency of their occurrence, which may vary from one setting to another.", "author" : [ { "dropping-particle" : "", "family" : "Rigouts", "given" : "Leen", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Gumusboga", "given" : "Mourad", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rijk", "given" : "Willem Bram", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Nduwamahoro", "given" : "Elie", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Uwizeye", "given" : "C\u00e9cile", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jong", "given" : "Bouke", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Deun", "given" : "Armand", "non-dropping-particle" : "Van", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-1", "issue" : "8", "issued" : { "date-parts" : [ [ "2013", "8" ] ] }, "page" : "2641-5", "title" : "Rifampin resistance missed in automated liquid culture system for Mycobacterium tuberculosis isolates with specific rpoB mutations.", "type" : "article-journal", "volume" : "51" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=6e35a569-8c72-417e-ac7a-a65b1473fefd" ] } ], "mendeley" : { "formattedCitation" : "[6]", "plainTextFormattedCitation" : "[6]", "previouslyFormattedCitation" : "[6]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[6] All strains with genotypic resistance except one that was of borderline resistance, were correctly classed as resistant or highly resistant by RR. All would have been reported as susceptible by MGIT, if no egg based medium or molecular methodology had been available. Mutations in the rpoB were detected in all strains. The use of molecular methodologies for the early detection of resistance conferring mutations in primary specimens is endorsed by WHO.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1586/17476348.2.5.583", "ISSN" : "1747-6356", "PMID" : "20477293", "abstract" : "Growing concerns about the spread of multidrug-resistant tuberculosis (MDR-TB) and the emergence of extensively drug-resistant TB have triggered substantial interest in the development and application of rapid tests for the detection of drug-resistant TB. Molecular assays to detect gene mutations that signal drug resistance are widely recognized as being most suited for rapid diagnosis. Among molecular assays, line probe assays have shown great promise. Currently, two line probe assays are commercially available: the INNO-LiPA Rif. TB kit (Innogenetics NV, Gent, Belgium) and the GenoType MTBDRplus assay (Hain Lifescience GmbH, Nehren, Germany). Evidence from a systematic review suggests that INNO-LiPA is a highly sensitive and specific test for the detection of rifampicin resistance in culture isolates. The test, however, appeared to have relatively lower sensitivity when used directly on clinical specimens. Another meta-analysis showed that the GenoType MTBDR assays had excellent accuracy for rifampicin resistance, even when used directly on clinical specimens. While specificity was excellent for isoniazid, sensitivity estimates were modest and variable. Based on evidence and expert opinion, the WHO recently endorsed the use of molecular line probe assays for rapid screening of patients at risk of MDR-TB. Special initiatives have been announced to make these assays accessible and affordable for countries with high MDR-TB prevalence. With strong evidence and new policy directives, the stage is now set for the use of rapid tests for MDR-TB diagnosis. Whether molecular tools, such as line probe assays, will actually make a clinical and public-health impact remains to be determined.", "author" : [ { "dropping-particle" : "", "family" : "Ling", "given" : "Daphne I", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Zwerling", "given" : "Alice A", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Pai", "given" : "Madhukar", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Expert review of respiratory medicine", "id" : "ITEM-1", "issue" : "5", "issued" : { "date-parts" : [ [ "2008", "10" ] ] }, "page" : "583-8", "title" : "Rapid diagnosis of drug-resistant TB using line probe assays: from evidence to policy.", "type" : "article-journal", "volume" : "2" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=7bfe7b9b-f546-404e-929f-4ea60736c3b8" ] }, { "id" : "ITEM-2", "itemData" : { "URL" : "http://www.ncbi.nlm.nih.gov/pubmed/26158191", "accessed" : { "date-parts" : [ [ "2015", "11", "9" ] ] }, "id" : "ITEM-2", "issued" : { "date-parts" : [ [ "0" ] ] }, "title" : "Policy Statement: Automated Real-Time Nucleic Acid Amplification Technology for Rapid and Simultaneous Detection of Tuberculosis and Rifampicin Res... - PubMed - NCBI", "type" : "webpage" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=4c069470-550e-4ce0-a34b-323fd187165f" ] } ], "mendeley" : { "formattedCitation" : "[14,15]", "plainTextFormattedCitation" : "[14,15]", "previouslyFormattedCitation" : "[14,15]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[14,15] The differentiation from non tuberculous mycobacteria in film positive samples and the detection of M. tuberculosis complex in smear negative samples is also a major advantage of these methods.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "ISSN" : "0125-2208", "PMID" : "25632618", "abstract" : "BACKGROUND: Incidence ofMDR-TB in Thailand is higher Diagnosis ofmulti-drug resistance (MDR-TB) is often delayed because of insufficient investigations. Therefore, there is a need to introduce an efficient testing method. Genotype MTBDR plus (Hain test) is a new technology of line probe assay (LPAs) In general practice, it had questionable benefit on negative and positive direct smear specimens.\n\nOBJECTIVE: Evaluate the accuracy of the Hain test as compared to the conventional culture and drug susceptibility test (DST), and evaluate the judgment of the physician on starting MDR-TB treatment after receiving the result of the Hain test.\n\nMATERIAL AND METHOD: An observational prospective study was done of 100 suspected MDR-TB patients who visited the Central Chest Institute of Thailand between September and December 2012. Ninety-four patients were included in the present study. They were assessed by direct sputum smear test, Hain test, sputum conventional culture, and drug-susceptibility test (DST) in the first visit. Followed-up treatment until cure/definite treatment were observed for definite diagnosis.\n\nRESULTS: Hain tests identified 65 (69%) MTB positives, 25 (26%) MTB negatives, andfour (4%) Non-tuberculous Mycobacteria (NTM), respectively. Fourteen of 17 negative direct smear but positive Hain test samples were culture negative. We followed these 14 patients. Four patients with TB treatment were cured. Six patients received MDR treatment and improved. Two patients were misdiagnosed CA. Two patients had old scar TB lesion. Sensitivity, specificity, and accuracy ofHain test for MDR-TB detection compared to standard conventional culture were 95%, 86%, and 88%, respectively. Twelve patients were NTMculture positive. Most of them (75%) were M abscessus. Sixty-three percent of chest physicians used the result of the Hain testfor extended treatment ofstandard regimen in non-IR resistance or changing ofTB regimen. There was correlation between judgments of starting MDR-TB treatment and results of Hain test (p = 0.001), no correlation between judgments of physician and pattern (default/relapse/failure) ofprevious TB treatment history (p = 0.6), and periods oftime from first diagnosis of TB to develop suspected MDR-TB (p = 0.09) were demonstrated.\n\nCONCLUSION: We recommend Hain test to diagnose MDR-TB in highly suspected cases, and to wait for conventional culture results to confirm NTM. Hain test has benefit in negative direct smear", "author" : [ { "dropping-particle" : "", "family" : "Sangsayunh", "given" : "Piamlarp", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Chuchothawon", "given" : "Chareon", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of the Medical Association of Thailand = Chotmaihet thangphaet", "id" : "ITEM-1", "issue" : "10", "issued" : { "date-parts" : [ [ "2014", "10" ] ] }, "page" : "1028-32", "title" : "Genotype MTBDR plus (Hain) test in suspected MDR-TB patients.", "type" : "article-journal", "volume" : "97" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=8e8b1744-4cdd-4df9-9f4f-f12ee7ee02c1" ] }, { "id" : "ITEM-2", "itemData" : { "DOI" : "10.1136/thoraxjnl-2015-207638", "ISSN" : "1468-3296", "PMID" : "27012887", "abstract" : "BACKGROUND: The quality of care for patients with TB in Eastern Europe has improved significantly; nevertheless drug resistance rates remain high. We analysed survival in a cohort of patients with multidrug-resistant and extensively drug-resistant (MDR-/XDR-) TB from Latvia, Lithuania, Estonia and Bucharest city.\n\nMETHODS: Consecutive adult new and retreatment patients with culture-confirmed pulmonary MDR-TB registered for treatment in 2009 (and in 2007 in Latvia) were enrolled; prospective survival information was collected.\n\nRESULTS: A total of 737 patients were included into the cohort. Of all MDR-TB cases, 46% were newly diagnosed; 56% of all MDR-TB cases had no additional resistance to fluoroquinolones or injectable agents, 33% had pre-XDR-TB and 11% XDR-TB. Median survival was 5.9\u2005years in patients with MDR-TB and XDR-TB; 1.9\u2005years in patients coinfected with HIV. Older age, male gender, alcohol abuse, retirement, co-morbidities, extrapulmonary involvement and HIV coinfection independently worsened survival. Inclusion of fluoroquinolones and injectable agents improves survival in patients with MDR-TB. Pre-XDR and XDR status did not significantly shorten survival as long as fluoroquinolones and injectable agents were part of the regimen. Moxifloxacin seems to improve survival in ofloxacin-susceptible patients when compared with older generation fluoroquinolones.\n\nCONCLUSIONS: The burden of additional resistances in patients with MDR-TB is high likely due to primary transmission of resistant strains. Social and programmatic factors including management of alcohol dependency, expansion of HIV testing and antiretroviral treatment need to be addressed in order to achieve cure and to interrupt transmission. The role of last generation fluoroquinolones and injectable agents in treatment of patients with pre-XDR and XDR-TB needs to be further investigated.", "author" : [ { "dropping-particle" : "", "family" : "Balabanova", "given" : "Yanina", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Ignatyeva", "given" : "Olga", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Fiebig", "given" : "Lena", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Riekstina", "given" : "Vija", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Danilovits", "given" : "Manfred", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jaama", "given" : "Kadri", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Davidaviciene", "given" : "Edita", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Radiulyte", "given" : "Birute", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Popa", "given" : "Christina Marcela", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Nikolayevskyy", "given" : "Vladyslav", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Drobniewski", "given" : "Francis", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Thorax", "id" : "ITEM-2", "issued" : { "date-parts" : [ [ "2016", "3", "24" ] ] }, "title" : "Survival of patients with multidrug-resistant TB in Eastern Europe: what makes a difference?", "type" : "article-journal" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=663a4f2d-1a03-4eca-95ca-1768c9c7fb7e" ] } ], "mendeley" : { "formattedCitation" : "[16,17]", "plainTextFormattedCitation" : "[16,17]", "previouslyFormattedCitation" : "[16,17]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[16,17] The introduction of molecular resistance detection as part of the routine assessment of tuberculosis samples would allow the detection of strains that are, as reported here, falsely susceptible by phenotypic liquid DST. After performing MIC determination in microtitre plates, tentative interpretation was performed based on the published data.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1007/s10096-011-1382-z", "ISSN" : "1435-4373", "PMID" : "21866324", "abstract" : "The agar proportion method (APM) for determining Mycobacterium tuberculosis susceptibilities is a qualitative method that requires 21 days in order to produce the results. The Sensititre method allows for a quantitative assessment. Our objective was to compare the accuracy, time to results, and ease of use of the Sensititre method to the APM. 7H10 plates in the APM and 96-well microtiter dry MYCOTB panels containing 12 antibiotics at full dilution ranges in the Sensititre method were inoculated with M. tuberculosis and read for colony growth. Thirty-seven clinical isolates were tested using both methods and 26 challenge strains of blinded susceptibilities were tested using the Sensititre method only. The Sensititre method displayed 99.3% concordance with the APM. The APM provided reliable results on day 21, whereas the Sensititre method displayed consistent results by day 10. The Sensititre method provides a more rapid, quantitative, and efficient method of testing both first- and second-line drugs when compared to the gold standard. It will give clinicians a sense of the degree of susceptibility, thus, guiding the therapeutic decision-making process. Furthermore, the microwell plate format without the need for instrumentation will allow its use in resource-poor settings.", "author" : [ { "dropping-particle" : "", "family" : "Abuali", "given" : "M M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Katariwala", "given" : "R", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "LaBombardi", "given" : "V J", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology", "id" : "ITEM-1", "issue" : "5", "issued" : { "date-parts" : [ [ "2012", "5" ] ] }, "page" : "835-9", "title" : "A comparison of the Sensititre\u00ae MYCOTB panel and the agar proportion method for the susceptibility testing of Mycobacterium tuberculosis.", "type" : "article-journal", "volume" : "31" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=fec311e6-682e-49e7-a0ba-1e7ca76bf561" ] }, { "id" : "ITEM-2", "itemData" : { "DOI" : "10.1128/JCM.02048-12", "ISSN" : "1098-660X", "PMID" : "22895034", "abstract" : "The Sensititre MycoTB plate (TREK Diagnostic Systems, Cleveland, OH) uses a microtiter plate MIC format for susceptibility testing of Mycobacterium tuberculosis complex isolates against first- and second-line antituberculosis agents. Categorical agreement versus the agar proportion method for 122 M. tuberculosis complex isolates was 94% to 100%.", "author" : [ { "dropping-particle" : "", "family" : "Hall", "given" : "Leslie", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jude", "given" : "Kurt P", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Clark", "given" : "Shirley L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Dionne", "given" : "Kim", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Merson", "given" : "Ryan", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Boyer", "given" : "Ana", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Parrish", "given" : "Nicole M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Wengenack", "given" : "Nancy L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-2", "issue" : "11", "issued" : { "date-parts" : [ [ "2012", "11" ] ] }, "page" : "3732-4", "title" : "Evaluation of the Sensititre MycoTB plate for susceptibility testing of the Mycobacterium tuberculosis complex against first- and second-line agents.", "type" : "article-journal", "volume" : "50" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=968c8c2a-2773-491e-a29a-fa678555299f" ] }, { "id" : "ITEM-3", "itemData" : { "DOI" : "10.1128/AAC.01209-13", "ISSN" : "1098-6596", "PMID" : "24100497", "abstract" : "For Mycobacterium tuberculosis, phenotypic methods for drug susceptibility testing of second-line drugs are poorly standardized and technically challenging. The Sensititre MYCOTB MIC plate (MYCOTB) is a microtiter plate containing lyophilized antibiotics and configured for determination of MICs to first- and second-line antituberculosis drugs. To evaluate the performance of MYCOTB for M. tuberculosis drug susceptibility testing using the Middlebrook 7H10 agar proportion method (APM) as the comparator, we conducted a two-site study using archived M. tuberculosis isolates from Uganda and the Republic of Korea. Thawed isolates were subcultured, and dilutions were inoculated into MYCOTB wells and onto 7H10 agar. MYCOTB results were read at days 7, 10, 14, and 21; APM results were read at 21 days. A total of 222 isolates provided results on both platforms. By APM, 106/222 (47.7%) of isolates were resistant to at least isoniazid and rifampin. Agreement between MYCOTB and APM with respect to susceptibility or resistance was \u226592% for 7 of 12 drugs when a strict definition was used and \u226596% for 10 of 12 drugs when agreement was defined by allowing a \u00b1 one-well range of dilutions around the APM critical concentration. For ethambutol, agreement was 80% to 81%. For moxifloxacin, agreement was 83% to 85%; incorporating existing DNA sequencing information for discrepant analysis raised agreement to 91% to 96%. For MYCOTB, the median time to plate interpretation was 10 days and interreader agreement was \u226595% for all drugs. MYCOTB provided reliable results for M. tuberculosis susceptibility testing of first- and second-line drugs except ethambutol, and results were available sooner than those determined by APM.", "author" : [ { "dropping-particle" : "", "family" : "Lee", "given" : "Jongseok", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Armstrong", "given" : "Derek T", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Ssengooba", "given" : "Willy", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Park", "given" : "Jeong-Ae", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Yu", "given" : "Yeuni", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Mumbowa", "given" : "Francis", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Namaganda", "given" : "Carolyn", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Mboowa", "given" : "Gerald", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Nakayita", "given" : "Germine", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Armakovitch", "given" : "Sandra", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Chien", "given" : "Gina", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Cho", "given" : "Sang-Nae", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Via", "given" : "Laura E", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Barry", "given" : "Clifton E", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Ellner", "given" : "Jerrold J", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Alland", "given" : "David", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Dorman", "given" : "Susan E", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Joloba", "given" : "Moses L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Antimicrobial agents and chemotherapy", "id" : "ITEM-3", "issue" : "1", "issued" : { "date-parts" : [ [ "2014", "1" ] ] }, "page" : "11-8", "title" : "Sensititre MYCOTB MIC plate for testing Mycobacterium tuberculosis susceptibility to first- and second-line drugs.", "type" : "article-journal", "volume" : "58" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=4092c78b-0ad4-4cb8-b848-d6d37fc83183" ] } ], "mendeley" : { "formattedCitation" : "[10,13,18]", "plainTextFormattedCitation" : "[10,13,18]", "previouslyFormattedCitation" : "[10,13,18]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[10,13,18] These publications applied the CLSI agar proportion method (APM) critical concentration to categorise the MIC values into resistant or susceptible. For rifampicin, resistance is defined as an MIC >1.0 mg/L. Interestingly 7 of the strains gave a rifampicin MIC of 1.0 mg/L which is strictly susceptible by APM breakpoint criteria, though very close to the cut-off. All the other strains had a rifampicin MIC greater than 1.0 mg/L. This may indicate that the current cut off is inappropriate for the detection of resistance associated with these mutations. In other work by two of the authors using the sensititre microdilution plates for MIC testing they found that all rifampicin susceptible strains had rifampicin MIC values of 0.25 mg/L or below (unpublished data). Three recent WHO External Quality Control distributions have not included any strains with an MIC of 1.0 mg/L or below and recent review of 44 results from 38 patients with mono, multi or extremely drug resistant isolates did not find this MIC, so it may not be common. (PC and IFL unpublished data) Based on those findings we suggest that resistance by this method be defined as >0.5mg/L. In egg based media, the drug concentrations tested are not directly related to actual MICs since the degradation and absorption of antimicrobials is high. Hence, the drug concentrations used are significantly higher than those used in Middlebrook media based methodologies,ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1093/jac/dkf214", "ISSN" : "14602091", "abstract" : "The concentrations of rifampicin in epithelial lining fluid (ELF), human alveolar macrophages (AM), bronchial mucosal biopsies and serum were measured after a single oral dose of 600 mg rifampicin from 15 patients undergoing fibre-optic bronchoscopy. The samples of ELF, AM, the bronchial mucosal biopsies and serum were obtained [~]2-5 h after dosing. The mean concentrations (mean {+/-} standard error) were 5.25 mg/L ({+/-}0.67) in ELF, 7.93 mg/kg ({+/-}1.61) in bronchial biopsies, 15.48 mg/L ({+/-}1.41) in serum and 251.8 mg/L ({+/-}65.92) in alveolar macrophages. These site concentrations exceeded the MIC90 for common respiratory pathogens including Mycobacterium tuberculosis and support the observed clinical efficacy of rifampicin in the treatment of a wide range of respiratory tract infections.", "author" : [ { "dropping-particle" : "", "family" : "Ziglam", "given" : "H. M.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of Antimicrobial Chemotherapy", "id" : "ITEM-1", "issue" : "6", "issued" : { "date-parts" : [ [ "2002", "11", "18" ] ] }, "page" : "1011-1015", "title" : "Rifampicin concentrations in bronchial mucosa, epithelial lining fluid, alveolar macrophages and serum following a single 600 mg oral dose in patients undergoing fibre-optic bronchoscopy", "type" : "article-journal", "volume" : "50" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=69f57b16-ff5a-4703-ab8e-3aae4208d824" ] } ], "mendeley" : { "formattedCitation" : "[19]", "plainTextFormattedCitation" : "[19]", "previouslyFormattedCitation" : "[19]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[19] in which there is a closer relation with actual MIC.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "ISBN" : "0849367166", "abstract" : "This book is comprised of reviews on the chemotherapy of mycobacterial infections, as well as descriptions of established methods and new techniques for drug susceptibility testing. Some of the fascinating topics examined include the activity of conventional and experimental antimicrobial agents, the rationale of drug combinations in chemotherapy, pharmacokinetics, and the problems of drug susceptibility of mycobacteria analyzed using standards established in other fields of clinical microbiology. Any physician or researcher involved with the therapy of tuberculosis, leprosy, M. avium in AIDS patients and other mycobacterial infections, and drug susceptibility testing will discover a wealth of information in this comprehensive volume.", "author" : [ { "dropping-particle" : "", "family" : "Heifets", "given" : "Leonid B.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "1991" ] ] }, "number-of-pages" : "224", "publisher" : "CRC Press", "title" : "Drug Susceptibility in the Chemotherapy of Mycobacterial Infections", "type" : "book" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=85773d74-4014-4fde-85d6-3530b7c40130" ] } ], "mendeley" : { "formattedCitation" : "[20]", "plainTextFormattedCitation" : "[20]", "previouslyFormattedCitation" : "[20]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[20] However, the addition of agar to Middlebrook media created problems as well due to protein binding and decay of the drugs related to prolonged incubation.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "ISBN" : "0781730147", "abstract" : "Long considered the definitive work in its field, this new edition presents all the principles and practices students need for a solid grounding in all aspects of clinical microbiology bacteriology, mycology, parasitology, and virology. Tests are presented according to the Clinical and Laboratory Standards Institute (formerly NCCLS) format. Easy-to-read charts outline theory, procedure, and interpretation of tests. Clinical correlations link microorganisms to specific disease states. Over 600 color plates depict the salient identification features of organisms. Numerous drawings, photographs, and tables clarify complex concepts.", "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "2006" ] ] }, "number-of-pages" : "1565", "publisher" : "Lippincott Williams & Wilkins", "title" : "Koneman's Color Atlas and Textbook of Diagnostic Microbiology", "type" : "book" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=6038fb1e-7b70-468d-a75b-9b14aa7196cd" ] } ], "mendeley" : { "formattedCitation" : "[21]", "plainTextFormattedCitation" : "[21]", "previouslyFormattedCitation" : "[21]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[21] Nevertheless, until recent reports of the phenomenon discussed in this article, correlation between DST in solid medium and MGIT for rifampicin had been reported to be excellent.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1128/JCM.44.3.811-818.2006", "ISBN" : "0095-1137 (Print)\\r0095-1137 (Linking)", "ISSN" : "00951137", "PMID" : "16517859", "abstract" : "The objectives of this study were to (i) compare agreement of the MGIT 960 system for first-line drugs with a methodology (the resistance ratio method [RRM]) that had been used in clinical trials, relating drug susceptibility to clinical outcome; (ii) compare the performance of the MGIT 960, RRM, and microtiter plate assay (MPA) methodologies for second-line drug testing; and (iii) define critical concentrations for ciprofloxacin and moxifloxacin for liquid-culture-based testing. The large collection of clinical isolates of Mycobacterium tuberculosis (n = 247) used included 176 (71%) multidrug-resistant isolates. The results for MGIT 960 and the RRM for rifampin and isoniazid (n = 200) were in excellent (99 to 100%) agreement for all strains. For streptomycin, 97% of the results at the critical concentration and 92% at high concentration, and for pyrazinamide 92% of results overall, were concordant, but for ethambutol, fewer than 85% (65% for the critical concentration and 84% for the high concentration) of the MGIT-based results were concordant with those for the RRM. The MGIT 960, RRM, and MPA assays (n = 133) correlated well for most second-line drugs tested. For susceptibility to ofloxacin, the MGIT 960 and MPA results were in full agreement. The amikacin and rifabutin results obtained by MGIT 960 agreed with the RRM results in 131 (99%) cases, and for capreomycin, they agreed for 129 of 133 isolates tested (97%). For prothionamide testing, only a limited number of drug-resistant isolates were available for testing and drawing definitive conclusions. We propose critical concentrations of 1.0 microg/ml and 0.125 microg/ml for ciprofloxacin and moxifloxacin, respectively, for liquid-culture-based testing.", "author" : [ { "dropping-particle" : "", "family" : "Kr\u00fc\u00fcner", "given" : "Annika", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Yates", "given" : "Malcolm D.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Drobniewski", "given" : "Francis a.", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of Clinical Microbiology", "id" : "ITEM-1", "issue" : "3", "issued" : { "date-parts" : [ [ "2006", "3" ] ] }, "page" : "811-818", "title" : "Evaluation of MGIT 960-based antimicrobial testing and determination of critical concentrations of first- and second-line antimicrobial drugs with drug-resistant clinical strains of Mycobacterium tuberculosis", "type" : "article-journal", "volume" : "44" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=5c3c4be2-b71b-4f31-bb6a-e68d6f6730cf" ] } ], "mendeley" : { "formattedCitation" : "[22]", "plainTextFormattedCitation" : "[22]", "previouslyFormattedCitation" : "[22]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[22] Following current recommendations for reading the proportion method in Middlebrook 7H10 plates, all save one would have been misidentified as susceptible.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "author" : [ { "dropping-particle" : "", "family" : "M-", "given" : "Clsi", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Desmond", "given" : "Edward", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Brown-elliott", "given" : "Barbara", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "id" : "ITEM-1", "issue" : "June", "issued" : { "date-parts" : [ [ "2011" ] ] }, "page" : "1-87", "title" : "Susceptibility Testing of Mycobacteria , , and Other Aerobic Actinomycetes y M24- -A2 Authors", "type" : "article-journal" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=b8ff9443-92a8-44a9-bdd7-07c77a2a422b" ] } ], "mendeley" : { "formattedCitation" : "[23]", "plainTextFormattedCitation" : "[23]", "previouslyFormattedCitation" : "[23]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[23] Bearing in mind that Middlebrook 7H9 and 7H10 only differ in the presence of agar and that both failed to identify rifampicin resistance, the problem could be one of media or that the cut off of 1 is too high to detect all truly resistant strains. Rifampicin decay occurs in both LJ and Middlebrook. In the former, it happens during inspissation in addition to the degradation that takes place during incubation so, it remains unclear why Middlebook medium fails to detect rifampicin resistance.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1128/JCM.01951-10", "ISSN" : "1098-660X", "PMID" : "21177895", "abstract" : "The objectives of this study were to monitor the stability of rifampin (RIF) in L\u00f6wenstein-Jensen medium (L-J medium) and 7H9 broth, which are the media commonly used for drug susceptibility testing (DST) of Mycobacterium tuberculosis. Rifampin degradation in stock solution, 7H9 broth, and L-J medium and during the inspissation process for L-J medium preparation was serially monitored by high-performance liquid chromatography (HPLC). L-J medium-based DST was conducted to examine the effect of L-J medium storage on the DST outcome. The RIF stock solution was stable for at least 3 months when kept at either 4\u00b0C or -20\u00b0C; RIF in 7H9 broth and L-J medium was almost 50% decayed after 1 week of storage at 37\u00b0C, and rifampin could not be detected in 7H9 or L-J medium after 3 weeks or 6 weeks of storage at 37\u00b0C. Approximately half of the drug was decomposed after 4 months of storage at 4\u00b0C for both media, and after 6 months of storage at 4\u00b0C, RIF in L-J medium was undetectable, while 38% of RIF remained in 7H9 medium. Approximately 21, 24, 29, and 35% RIF degradations were detected when the L-J medium was coagulated at 75\u00b0C, 80\u00b0C, 85\u00b0C, and 90\u00b0C, respectively. The DST outcomes when using L-J medium stored for different periods of time were consistent with the RIF concentration monitoring data. Rifampin in stock solution is stable for at least 3 months at a reduced storage temperature. Media containing RIF should be prepared strictly according to validated standard operating procedures. RIF degradation is a possible reason for false resistance categorizations of M. tuberculosis isolates in the clinical laboratory.", "author" : [ { "dropping-particle" : "", "family" : "Yu", "given" : "Xia", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jiang", "given" : "Guanglu", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Li", "given" : "Huiwen", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Zhao", "given" : "Yanlin", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Zhang", "given" : "Hongtao", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Zhao", "given" : "Liping", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Ma", "given" : "Yu", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Coulter", "given" : "Christopher", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Huang", "given" : "Hairong", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-1", "issue" : "3", "issued" : { "date-parts" : [ [ "2011", "3", "1" ] ] }, "language" : "en", "page" : "784-9", "publisher" : "American Society for Microbiology (ASM)", "title" : "Rifampin stability in 7H9 broth and L\u00f6wenstein-Jensen medium.", "type" : "article-journal", "volume" : "49" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=979ee1d4-bf28-4f94-97bc-d8e5272c78ce" ] } ], "mendeley" : { "formattedCitation" : "[24]", "plainTextFormattedCitation" : "[24]", "previouslyFormattedCitation" : "[24]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[24] Following the original report of the MGIT missing rifampicin resistance,ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1128/JCM.02741-12", "ISSN" : "1098-660X", "PMID" : "23761146", "abstract" : "WHO-endorsed phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are assumed to be the gold standard for identifying rifampin (RMP) resistance. However, previous results indicated that low-level, yet probably clinically relevant, RMP resistance linked to specific rpoB mutations is easily missed by some growth-based methods. We aimed to compare the level of resistance detected on L\u00f6wenstein-Jensen (LJ) medium with resistance detected by the Bactec MGIT 960 automated DST (MGIT-DST) system for various rpoB mutants. Full agreement between LJ and MGIT-DST was observed for mutations located at codons 513 (Lys or Pro) and 531 (Leu, Trp), which were always resistant by both methods. For mutations 511Pro, 516Tyr, 533Pro, 572Phe, and several 526 mutations, LJ and MGIT results were highly discordant, with MGIT-DST failing to give a result or declaring the strains susceptible. Our data show that phenotypic RMP resistance testing of M. tuberculosis is not a binary phenomenon for some rpoB mutations and that the widely used automated MGIT 960 system is prone to miss some RMP resistance-conferring mutations, while careful DST on LJ missed hardly any. Given the association of these mutations with poor clinical outcome, our findings suggest that the gold standard for rifampin resistance should be reconsidered, in order to address the present confusion caused by discrepancies between phenotypic and genotypic results. The impacts of these mutations will depend on the frequency of their occurrence, which may vary from one setting to another.", "author" : [ { "dropping-particle" : "", "family" : "Rigouts", "given" : "Leen", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Gumusboga", "given" : "Mourad", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rijk", "given" : "Willem Bram", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Nduwamahoro", "given" : "Elie", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Uwizeye", "given" : "C\u00e9cile", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jong", "given" : "Bouke", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Deun", "given" : "Armand", "non-dropping-particle" : "Van", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-1", "issue" : "8", "issued" : { "date-parts" : [ [ "2013", "8" ] ] }, "page" : "2641-5", "title" : "Rifampin resistance missed in automated liquid culture system for Mycobacterium tuberculosis isolates with specific rpoB mutations.", "type" : "article-journal", "volume" : "51" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=6e35a569-8c72-417e-ac7a-a65b1473fefd" ] } ], "mendeley" : { "formattedCitation" : "[6]", "plainTextFormattedCitation" : "[6]", "previouslyFormattedCitation" : "[6]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[6] it transpired that this is not just an in vitro laboratory problem but that clinical failure follows when patients carrying strains with this pheno/genotype are treated with standard rifampicin-based regimens.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1371/journal.pone.0090569", "ISSN" : "1932-6203", "PMID" : "24599230", "abstract" : "The World Health Organization has recommended use of molecular-based tests MTBDRplus and GeneXpert MTB/RIF to diagnose multidrug-resistant tuberculosis in developing and high-burden countries. Both tests are based on detection of mutations in the Rifampin (RIF) Resistance-Determining Region of DNA-dependent RNA Polymerase gene (rpoB). Such mutations are found in 95-98% of Mycobacterium tuberculosis strains determined to be RIF-resistant by the \"gold standard\" culture-based drug susceptibility testing (DST). We report the phenotypic and genotypic characterization of 153 consecutive clinical Mycobacterium tuberculosis strains diagnosed as RIF-resistant by molecular tests in our laboratory in Port-au-Prince, Haiti. 133 isolates (86.9%) were resistant to both RIF and Isoniazid and 4 isolates (2.6%) were RIF mono-resistant in MGIT SIRE liquid culture-based DST. However the remaining 16 isolates (10.5%) tested RIF-sensitive by the assay. Five strains with discordant genotypic and phenotypic susceptibility results had RIF minimal inhibitory concentration (MIC) close to the cut-off value of 1 \u00b5g/ml used in phenotypic susceptibility assays and were confirmed as resistant by DST on solid media. Nine strains had sub-critical RIF MICs ranging from 0.063 to 0.5 \u00b5g/ml. Finally two strains were pan-susceptible and harbored a silent rpoB mutation. Our data indicate that not only detection of the presence but also identification of the nature of rpoB mutation is needed to accurately diagnose resistance to RIF in Mycobacterium tuberculosis. Observed clinical significance of low-level resistance to RIF supports the re-evaluation of the present critical concentration of the drug used in culture-based DST assays.", "author" : [ { "dropping-particle" : "", "family" : "Ocheretina", "given" : "Oksana", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Escuyer", "given" : "Vincent E", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Mabou", "given" : "Marie-Marcelle", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Royal-Mardi", "given" : "Gertrude", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Collins", "given" : "Sean", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Vilbrun", "given" : "Stalz C", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Pape", "given" : "Jean W", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Fitzgerald", "given" : "Daniel W", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "PloS one", "id" : "ITEM-1", "issue" : "3", "issued" : { "date-parts" : [ [ "2014", "1", "5" ] ] }, "page" : "e90569", "publisher" : "Public Library of Science", "title" : "Correlation between genotypic and phenotypic testing for resistance to rifampin in Mycobacterium tuberculosis clinical isolates in Haiti: investigation of cases with discrepant susceptibility results.", "type" : "article-journal", "volume" : "9" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=3e72aa60-839f-4376-88fc-a06f38797798" ] }, { "id" : "ITEM-2", "itemData" : { "DOI" : "10.1128/JCM.00553-13", "ISSN" : "1098-660X", "PMID" : "23761144", "abstract" : "The rapid diagnosis of rifampin resistance is hampered by a reported insufficient specificity of molecular techniques for detection of rpoB mutations. Our objective for this study was to document the prevalence and prognostic value of rpoB mutations with unclear phenotypic resistance. The study design entailed sequencing directly from sputum of first failure or relapse patients without phenotypic selection and comparison of the standard retreatment regimen outcome, according to the mutation present. We found that among all rpoB mutations, the best-documented \"disputed\" rifampin resistance mutations (511Pro, 516Tyr, 526Asn, 526Leu, 533Pro, and 572Phe) made up 13.1% and 10.6% of all mutations in strains from Bangladesh and Kinshasa, respectively. Except for the 511Pro and 526Asn mutations, most of these strains with disputed mutations tested rifampin resistant in routine L\u00f6wenstein-Jensen medium proportion method drug susceptibility testing (DST; 78.7%), but significantly less than those with common, undisputed mutations (96.3%). With 63% of patients experiencing failure or relapse in both groups, there was no difference in outcome of first-line retreatment between patients carrying a strain with disputed versus common mutations. We conclude that rifampin resistance that is difficult to detect by the gold standard, phenotypic DST, is clinically and epidemiologically highly relevant. Sensitivity rather than specificity is imperfect with any rifampin DST method. Even at a low prevalence of rifampin resistance, a rifampin-resistant result issued by a competent laboratory may not warrant confirmation, although the absence of a necessity for confirmation needs to be confirmed for molecular results among new cases. However, a result of rifampin susceptibility should be questioned when suspicion is very high, and further DST using a different system (i.e., genotypic after phenotypic testing) would be fully justified.", "author" : [ { "dropping-particle" : "", "family" : "Deun", "given" : "Armand", "non-dropping-particle" : "Van", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Aung", "given" : "Kya J M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Bola", "given" : "Valentin", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Lebeke", "given" : "Rossin", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Hossain", "given" : "Mohamed Anwar", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rijk", "given" : "Willem Bram", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rigouts", "given" : "Leen", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Gumusboga", "given" : "Aysel", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Torrea", "given" : "Gabriela", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jong", "given" : "Bouke C", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-2", "issue" : "8", "issued" : { "date-parts" : [ [ "2013", "8" ] ] }, "page" : "2633-40", "title" : "Rifampin drug resistance tests for tuberculosis: challenging the gold standard.", "type" : "article-journal", "volume" : "51" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=0258cb93-a441-4941-b259-883578d2f7db" ] }, { "id" : "ITEM-3", "itemData" : { "DOI" : "10.5588/ijtld.14.0651", "ISSN" : "1815-7920", "PMID" : "25574917", "abstract" : "SETTING: Greater Mymensingh area, Bangladesh.\n\nOBJECTIVES: To document among new tuberculosis (TB) patients the proportions and treatment outcomes of silent, non-disputed and disputed (generally missed by rapid drug susceptibility testing [DST]) rpoB mutations, and their detection by commercial molecular assays.\n\nDESIGN: Retrospective analysis of rpoB sequences from randomly selected ethanol-preserved diagnostic sputum samples; comparison of sequencing with conventional DST results and standard first-line treatment outcome; retesting of samples with mutations using the Xpert MTB/RIF and GenoType MTBDRplus assays.\n\nRESULTS: Of 1091 samples, 5.8% failed amplification, and six contained other mycobacteria. In 2005 and 2010, respectively 2/500 (0.4%) and 11/522 (2.1%) amplicons showed non-silent mutations. At least 7/13 of these belonged to the disputed group, with 5/7 patients suffering adverse treatment outcome. One silent mutation went undetected by commercial assays. Following routine DST indications, only three cases with a non-silent mutation were eventually detected.\n\nCONCLUSIONS: Disputed rpoB mutations may be responsible for the majority of rifampicin (RMP) resistance among new cases, and lead to adverse outcomes of first-line treatment. Silent mutations do not necessarily cause Xpert or line-probe assay false RMP-resistant results. Molecular RMP DST could greatly simplify resistance surveillance, in addition to offering the best prospects for early and accurate individual diagnosis.", "author" : [ { "dropping-particle" : "", "family" : "Deun", "given" : "A", "non-dropping-particle" : "Van", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Aung", "given" : "K J M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Hossain", "given" : "A", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rijk", "given" : "P", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Gumusboga", "given" : "M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rigouts", "given" : "L", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jong", "given" : "B C", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" } ], "container-title" : "The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease", "id" : "ITEM-3", "issue" : "2", "issued" : { "date-parts" : [ [ "2015", "2" ] ] }, "page" : "185-90", "title" : "Disputed rpoB mutations can frequently cause important rifampicin resistance among new tuberculosis patients.", "type" : "article-journal", "volume" : "19" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=6e1dd858-9c07-4eac-a307-429db96f9ea8" ] } ], "mendeley" : { "formattedCitation" : "[25\u201327]", "plainTextFormattedCitation" : "[25\u201327]", "previouslyFormattedCitation" : "[25\u201327]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[2527] Mutations at 516 and 526 have been associated with resistance to rifampicin and susceptibility to rifabutin. Mutations H526L, D516Y and L511P confer a low level of resistance, as previously reported.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1093/jac/dkp204", "ISSN" : "1460-2091", "PMID" : "19520715", "abstract" : "OBJECTIVES: The aim of this study was to compare the efficiency of two commercial assays, INNO-LiPA Rif.TB and MTBDRplus, for the identification of mutations in the rpoB hot-spot region and to assess the efficiency of these mutations in conferring resistance to rifampicin.\n\nMETHODS: A collection of 126 rifampicin-resistant Mycobacterium tuberculosis and Mycobacterium africanum isolates and 18 rifampicin-susceptible isolates from different countries were analysed using the two hybridization assays.\n\nRESULTS: For 60 strains the hot-spot region of the rpoB gene was sequenced, confirming the results of the hybridization assays and allowing the identification of new mutations. In total, 17 mutations involving 10 codons were observed, two of which are newly described (D516Y and E562G/P564L). Mutations L533P, H526L, D516Y and L511P and the double mutation E562G/P564L conferred a low level of resistance.\n\nCONCLUSIONS: The assays INNO-LiPA Rif.TB and MTBDRplus identified rpoB mutations in 98.4% of cases. MTBDRplus provided additional information due to the overlapping hybridization probes and in addition allowed the investigation of katG mutations for isoniazid resistance.", "author" : [ { "dropping-particle" : "", "family" : "Hauck", "given" : "Yolande", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Fabre", "given" : "Michel", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Vergnaud", "given" : "Gilles", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Soler", "given" : "Charles", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Pourcel", "given" : "Christine", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "container-title" : "The Journal of antimicrobial chemotherapy", "id" : "ITEM-1", "issue" : "2", "issued" : { "date-parts" : [ [ "2009", "8", "1" ] ] }, "page" : "259-62", "title" : "Comparison of two commercial assays for the characterization of rpoB mutations in Mycobacterium tuberculosis and description of new mutations conferring weak resistance to rifampicin.", "type" : "article-journal", "volume" : "64" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=d5971db6-9a38-410c-8b7d-837a8fd31d4d" ] } ], "mendeley" : { "formattedCitation" : "[28]", "plainTextFormattedCitation" : "[28]", "previouslyFormattedCitation" : "[28]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[28] These mutations made up 13.1% and 10.6% of all mutations in strains from Bangladesh and Kinshasa.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "DOI" : "10.1128/JCM.00553-13", "ISSN" : "1098-660X", "PMID" : "23761144", "abstract" : "The rapid diagnosis of rifampin resistance is hampered by a reported insufficient specificity of molecular techniques for detection of rpoB mutations. Our objective for this study was to document the prevalence and prognostic value of rpoB mutations with unclear phenotypic resistance. The study design entailed sequencing directly from sputum of first failure or relapse patients without phenotypic selection and comparison of the standard retreatment regimen outcome, according to the mutation present. We found that among all rpoB mutations, the best-documented \"disputed\" rifampin resistance mutations (511Pro, 516Tyr, 526Asn, 526Leu, 533Pro, and 572Phe) made up 13.1% and 10.6% of all mutations in strains from Bangladesh and Kinshasa, respectively. Except for the 511Pro and 526Asn mutations, most of these strains with disputed mutations tested rifampin resistant in routine L\u00f6wenstein-Jensen medium proportion method drug susceptibility testing (DST; 78.7%), but significantly less than those with common, undisputed mutations (96.3%). With 63% of patients experiencing failure or relapse in both groups, there was no difference in outcome of first-line retreatment between patients carrying a strain with disputed versus common mutations. We conclude that rifampin resistance that is difficult to detect by the gold standard, phenotypic DST, is clinically and epidemiologically highly relevant. Sensitivity rather than specificity is imperfect with any rifampin DST method. Even at a low prevalence of rifampin resistance, a rifampin-resistant result issued by a competent laboratory may not warrant confirmation, although the absence of a necessity for confirmation needs to be confirmed for molecular results among new cases. However, a result of rifampin susceptibility should be questioned when suspicion is very high, and further DST using a different system (i.e., genotypic after phenotypic testing) would be fully justified.", "author" : [ { "dropping-particle" : "", "family" : "Deun", "given" : "Armand", "non-dropping-particle" : "Van", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Aung", "given" : "Kya J M", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Bola", "given" : "Valentin", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Lebeke", "given" : "Rossin", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Hossain", "given" : "Mohamed Anwar", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rijk", "given" : "Willem Bram", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Rigouts", "given" : "Leen", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Gumusboga", "given" : "Aysel", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Torrea", "given" : "Gabriela", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" }, { "dropping-particle" : "", "family" : "Jong", "given" : "Bouke C", "non-dropping-particle" : "de", "parse-names" : false, "suffix" : "" } ], "container-title" : "Journal of clinical microbiology", "id" : "ITEM-1", "issue" : "8", "issued" : { "date-parts" : [ [ "2013", "8" ] ] }, "page" : "2633-40", "title" : "Rifampin drug resistance tests for tuberculosis: challenging the gold standard.", "type" : "article-journal", "volume" : "51" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=0258cb93-a441-4941-b259-883578d2f7db" ] } ], "mendeley" : { "formattedCitation" : "[26]", "plainTextFormattedCitation" : "[26]", "previouslyFormattedCitation" : "[26]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[26] During the study period, 4.6% of TB cases in the UK were in patients born in Bangladesh in 2012 and 3.3% in 2013.ADDIN CSL_CITATION { "citationItems" : [ { "id" : "ITEM-1", "itemData" : { "author" : [ { "dropping-particle" : "", "family" : "PHE", "given" : "", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "id" : "ITEM-1", "issued" : { "date-parts" : [ [ "2013" ] ] }, "title" : "Tuberculosis in the UK: Annual report on tuberculosis surveillance in the UK, 2013", "type" : "article-journal" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=83d8e9bd-ac65-40f7-9e88-544c1dbe0b47" ] }, { "id" : "ITEM-2", "itemData" : { "author" : [ { "dropping-particle" : "", "family" : "PHE", "given" : "", "non-dropping-particle" : "", "parse-names" : false, "suffix" : "" } ], "id" : "ITEM-2", "issued" : { "date-parts" : [ [ "2014" ] ] }, "title" : "Tuberculosis in the UK: 2014 report", "type" : "article-journal" }, "uris" : [ "http://www.mendeley.com/documents/?uuid=c0ca024f-1f94-428b-90e6-6ba00bc2db83" ] } ], "mendeley" : { "formattedCitation" : "[29,30]", "plainTextFormattedCitation" : "[29,30]", "previouslyFormattedCitation" : "[29,30]" }, "properties" : { "noteIndex" : 0 }, "schema" : "https://github.com/citation-style-language/schema/raw/master/csl-citation.json" }[29,30] There appear to be 3 rpoB resistant genotypes: L511P+M515I seen only twice in this set, H526L seen once previously and D516Y seen on numerous occasions. Since no clusters were observed, it is likely that D516Y has been independently acquired in each case rather than it being present in a circulating clone. There are some limitations of this study. Some strains may have been missed since national reference centres receive samples/strains after they have been processed by regional and local laboratories. There is no clinical or patients outcome information associated to this strains so, outcome cannot be associated to any particular phenol/genotype. As observed previously, these phenol/genotypes have been found in certain countries, some of which are represented in the UK and Ireland. Unfortunately, in our study, patients country of origin is not always captured and even when it is, that information is not available for the laboratory. Conclusion: At present, knowledge about rarer mutations is not entirely clear so, phenotypic testing still provides invaluable information. Molecular DNA assays help to address the issue of the lack of an accurate and rapid method for detecting phenotypic resistance for MTB, but not all mutations confer phenotypic resistance. The accumulation of molecular data with correlating clinical outcome data will allow us in time to have a more thorough understanding of true drug resistance in MTB globally. Commercially available microtitre plates allow MIC determinations at a reasonable cost. However, in this study those plates using current cut-off misindentified 7 strains as susceptible. As stated above, 0.5mg/L could be a more accurate cut-off for all Middlebrook-based DST methodologies. The incidence of MGIT rifampicin sensitive, but RR rifampicin resistant isolates appears to be low in the UK and Ireland (0.23% of all processed strains and 5.26% of those that had second line DST done), but this may be an underestimate as some laboratories rely on MGIT alone to detect rifampicin resistance. Acknowledgements: We would like to thank colleagues for referring isolates and related information and to Mr Michael Smith for assisting with epidemiological data. We would like to thank Dr Michael Lockhart and other colleagues for referring isolates and related information and Mr Michael Smith for assisting with epidemiological data Prof Abubakar, Jennifer Davidson, Miranda Loutet and Maeve Lalor from PHE for assisting with the epidemiological data. Funding: This work is part of the remit of the SMRL funded through its contract with Health Protection Scotland and National Services Scotland. At the IRML and NMRL, this study was supported by internal funding. Transparency declarations: None to declare References ADDIN Mendeley Bibliography CSL_BIBLIOGRAPHY [1] Anuwatnonthakate A, Whitehead SJ, Varma JK, Silachamroon U, Kasetjaroen Y, Moolphate S, et al. Effect of mycobacterial drug resistance patterns on patients survival: a cohort study in Thailand. 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