Detection of P. malariae using a new rapid isothermal amplification lateral flow assay

P. malariae is found worldwide and causes chronic parasitism in its human hosts. We developed a P. malariae (Pm) diagnostic assay that uses rapid, isothermal recombinase polymerase amplification (RPA) and lateral-flow-strip detection. Using 18S rRNA plasmid DNA, the assay demonstrates a detection li...

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Autores principales: Assefa, Ashenafi, Wamae, Kevin K., Hennelly, Chris M., Ngasala, Billy, Muller, Meredith, Kalonji, Albert, Phanzu, Fernandine, Cunningham, Clark H., Lin, Jessica T., Parr, Jonathan B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10543045/
https://www.ncbi.nlm.nih.gov/pubmed/37790493
http://dx.doi.org/10.1101/2023.02.26.23286371
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author Assefa, Ashenafi
Wamae, Kevin K.
Hennelly, Chris M.
Ngasala, Billy
Muller, Meredith
Kalonji, Albert
Phanzu, Fernandine
Cunningham, Clark H.
Lin, Jessica T.
Parr, Jonathan B.
author_facet Assefa, Ashenafi
Wamae, Kevin K.
Hennelly, Chris M.
Ngasala, Billy
Muller, Meredith
Kalonji, Albert
Phanzu, Fernandine
Cunningham, Clark H.
Lin, Jessica T.
Parr, Jonathan B.
author_sort Assefa, Ashenafi
collection PubMed
description P. malariae is found worldwide and causes chronic parasitism in its human hosts. We developed a P. malariae (Pm) diagnostic assay that uses rapid, isothermal recombinase polymerase amplification (RPA) and lateral-flow-strip detection. Using 18S rRNA plasmid DNA, the assay demonstrates a detection limit of 10 copies /μL (~1.7 genome equivalents) and 100% analytical specificity. Testing in field samples showed 95% clinical sensitivity and 88% specificity compared to qPCR. Total assay time was 35 minutes. Combined with simplified DNA extraction methods, the assay has potential for future field-deployable point-of-care use to detect a parasite species that remains largely undiagnosed.
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spelling pubmed-105430452023-10-03 Detection of P. malariae using a new rapid isothermal amplification lateral flow assay Assefa, Ashenafi Wamae, Kevin K. Hennelly, Chris M. Ngasala, Billy Muller, Meredith Kalonji, Albert Phanzu, Fernandine Cunningham, Clark H. Lin, Jessica T. Parr, Jonathan B. medRxiv Article P. malariae is found worldwide and causes chronic parasitism in its human hosts. We developed a P. malariae (Pm) diagnostic assay that uses rapid, isothermal recombinase polymerase amplification (RPA) and lateral-flow-strip detection. Using 18S rRNA plasmid DNA, the assay demonstrates a detection limit of 10 copies /μL (~1.7 genome equivalents) and 100% analytical specificity. Testing in field samples showed 95% clinical sensitivity and 88% specificity compared to qPCR. Total assay time was 35 minutes. Combined with simplified DNA extraction methods, the assay has potential for future field-deployable point-of-care use to detect a parasite species that remains largely undiagnosed. Cold Spring Harbor Laboratory 2023-03-01 /pmc/articles/PMC10543045/ /pubmed/37790493 http://dx.doi.org/10.1101/2023.02.26.23286371 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Assefa, Ashenafi
Wamae, Kevin K.
Hennelly, Chris M.
Ngasala, Billy
Muller, Meredith
Kalonji, Albert
Phanzu, Fernandine
Cunningham, Clark H.
Lin, Jessica T.
Parr, Jonathan B.
Detection of P. malariae using a new rapid isothermal amplification lateral flow assay
title Detection of P. malariae using a new rapid isothermal amplification lateral flow assay
title_full Detection of P. malariae using a new rapid isothermal amplification lateral flow assay
title_fullStr Detection of P. malariae using a new rapid isothermal amplification lateral flow assay
title_full_unstemmed Detection of P. malariae using a new rapid isothermal amplification lateral flow assay
title_short Detection of P. malariae using a new rapid isothermal amplification lateral flow assay
title_sort detection of p. malariae using a new rapid isothermal amplification lateral flow assay
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10543045/
https://www.ncbi.nlm.nih.gov/pubmed/37790493
http://dx.doi.org/10.1101/2023.02.26.23286371
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