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  4. Strain discrimination of Yersinia pestis using a SERS-based electrochemically driven melting curve analysis of variable number tandem repeat sequences
 
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Strain discrimination of Yersinia pestis using a SERS-based electrochemically driven melting curve analysis of variable number tandem repeat sequences
File(s)
c4sc03084b.pdf (1002.91 KB)
Published version
Author(s)
Papadopoulou, E
Gale, N
Goodchild, SA
Cleary, DW
Weller, SA
more
Type
Journal Article
Abstract
Strain discrimination within genetically highly similar bacteria is critical for epidemiological studies and forensic applications. An electrochemically driven melting curve analysis monitored by SERS has been utilised to reliably discriminate strains of the bacterial pathogen Yersinia pestis, the causative agent of plague. DNA amplicons containing Variable Number Tandem Repeats (VNTRs) were generated from three strains of Y. pestis: CO92, Harbin 35 and Kim. These amplicons contained a 10 base pair VNTR repeated 6, 5, and 4 times in CO92, Harbin 35 and Kim respectively. The assay also included a blocker oligonucleotide comprising 3 repeats of the 10-mer VNTR sequence. The use of the blocker reduced the effective length of the target sequence available to bind to the surface bound probe and significantly improved the sensitivity of the discrimination. The results were consistent during three replicates that were carried out on different days, using different batches of PCR product and different SERS sphere segment void (SSV) substrate. This methodology which combines low cost, speed and sensitivity is a promising alternative to the time consuming current electrophoretic methods.
Date Issued
2015-01-07
Date Acceptance
2014-12-23
Citation
Chemical Science, 2015, 6 (3), pp.1846-1852
URI
http://hdl.handle.net/10044/1/39903
DOI
https://www.dx.doi.org/10.1039/c4sc03084b
ISSN
2041-6520
Publisher
Royal Society of Chemistry
Start Page
1846
End Page
1852
Journal / Book Title
Chemical Science
Volume
6
Issue
3
Copyright Statement
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.
License URL
http://creativecommons.org/licenses/by/4.0/
Publication Status
Published
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