Sensitive detection of asymptomatic and symptomatic malaria with seven novel parasite-specific LAMP assays and translation for use at point-of-care
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Published version
Author(s)
Type
Journal Article
Abstract
Human malaria is a life-threatening parasitic disease with high impact in the sub-Saharan Africa region, where 95% of global cases occurred in 2021. While most malaria diagnostic tools are focused on Plasmodium falciparum, there is a current lack of testing non-P. falciparum cases, which may be underreported and, if undiagnosed or untreated, may lead to severe consequences. In this work, seven species-specific loop-mediated isothermal amplification (LAMP) assays were designed and evaluated against TaqMan quantitative PCR (qPCR), microscopy, and enzyme-linked immunosorbent assays (ELISAs). Their clinical performance was assessed with a cohort of 164 samples of symptomatic and asymptomatic patients from Ghana. All asymptomatic samples with a parasite load above 80 genomic DNA (gDNA) copies per μL of extracted sample were detected with the Plasmodium falciparum LAMP assay, reporting 95.6% (95% confidence interval [95% CI] of 89.9 to 98.5) sensitivity and 100% (95% CI of 87.2 to 100) specificity. This assay showed higher sensitivity than microscopy and ELISA, which were 52.7% (95% CI of 39.7 to 67%) and 67.3% (95% CI of 53.3 to 79.3%), respectively. Nine samples were positive for P. malariae, indicating coinfections with P. falciparum, which represented 5.5% of the tested population. No samples were detected as positive for P. vivax, P. ovale, P. knowlesi, or P. cynomolgi by any method. Furthermore, translation to the point-of-care was demonstrated with a subcohort of 18 samples tested locally in Ghana using our handheld lab-on-chip platform, Lacewing, showing comparable results to a conventional fluorescence-based instrument. The developed molecular diagnostic test could detect asymptomatic malaria cases, including submicroscopic parasitemia, and it has the potential to be used for point-of-care applications.
Date Issued
2023-05-09
Date Acceptance
2023-04-18
Citation
Microbiology Spectrum, 2023, 11 (3), pp.1-12
ISSN
2165-0497
Publisher
American Society for Microbiology
Start Page
1
End Page
12
Journal / Book Title
Microbiology Spectrum
Volume
11
Issue
3
Copyright Statement
© 2023 Malpartida-Cardenas et al.
This is an open-access article distributed under
the terms of the Creative Commons
Attribution 4.0 International license
This is an open-access article distributed under
the terms of the Creative Commons
Attribution 4.0 International license
License URL
Identifier
https://journals.asm.org/doi/10.1128/spectrum.05222-22
Publication Status
Published
Date Publish Online
2023-05-09