Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions

Copyright © 2015, American Society for Microbiology. All Rights Reserved. Flow cytometry is an objective method for conducting in vitro antimalarial sensitivity assays with increasing potential for application in field sites. We examined in vitro susceptibility to seven anti-malarial drugs for 40 fr...

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Main Authors: Charles J. Woodrow, Chirapat Wangsing, Kanlaya Sriprawat, Peter R. Christensen, Francois Nosten, Laurent Rénia, Bruce Russell, Benoît Malleret
Other Authors: Nuffield Department of Clinical Medicine
Format: Article
Published: 2018
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Online Access:https://repository.li.mahidol.ac.th/handle/123456789/36314
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spelling th-mahidol.363142018-11-23T17:34:58Z Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions Charles J. Woodrow Chirapat Wangsing Kanlaya Sriprawat Peter R. Christensen Francois Nosten Laurent Rénia Bruce Russell Benoît Malleret Nuffield Department of Clinical Medicine Mahidol University Agency for Science, Technology and Research, Singapore Yong Loo Lin School of Medicine Medicine Copyright © 2015, American Society for Microbiology. All Rights Reserved. Flow cytometry is an objective method for conducting in vitro antimalarial sensitivity assays with increasing potential for application in field sites. We examined in vitro susceptibility to seven anti-malarial drugs for 40 fresh P. falciparum field isolates via a flow cytometry method (FCM), a colorimetric LDH-based ELISA (DELI), and standard microscopic slide analysis of growth. For FCM, 184/280 (66%) assays met analytical acceptance criteria, compared to 166/280 (59%) for DELI. There was good agreement between FCM and microscopy, while DELI tended to produce higher half-maximal inhibition constants (IC50s) than FCM, with an overall bias of 2.2-fold (Bland-Altman comparison). Values for artesunate and dihydroartemisinin were most affected. Paradoxical increases in signal at very high concentrations of mefloquine and related compounds were more marked with the DELI assay, suggesting that off-target effects on LDH production may be responsible. Loss of FCM signal due to reinvasion or slow growth was observed in a small number of samples. These results extend previous work on use of flow cytometry to determine antimalarial susceptibility in terms of the number of samples, range of drugs, and comparison with other methods. 2018-11-23T10:34:58Z 2018-11-23T10:34:58Z 2015-10-01 Article Journal of Clinical Microbiology. Vol.53, No.10 (2015), 3296-3303 10.1128/JCM.01226-15 1098660X 00951137 2-s2.0-84941949667 https://repository.li.mahidol.ac.th/handle/123456789/36314 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84941949667&origin=inward
institution Mahidol University
building Mahidol University Library
continent Asia
country Thailand
Thailand
content_provider Mahidol University Library
collection Mahidol University Institutional Repository
topic Medicine
spellingShingle Medicine
Charles J. Woodrow
Chirapat Wangsing
Kanlaya Sriprawat
Peter R. Christensen
Francois Nosten
Laurent Rénia
Bruce Russell
Benoît Malleret
Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions
description Copyright © 2015, American Society for Microbiology. All Rights Reserved. Flow cytometry is an objective method for conducting in vitro antimalarial sensitivity assays with increasing potential for application in field sites. We examined in vitro susceptibility to seven anti-malarial drugs for 40 fresh P. falciparum field isolates via a flow cytometry method (FCM), a colorimetric LDH-based ELISA (DELI), and standard microscopic slide analysis of growth. For FCM, 184/280 (66%) assays met analytical acceptance criteria, compared to 166/280 (59%) for DELI. There was good agreement between FCM and microscopy, while DELI tended to produce higher half-maximal inhibition constants (IC50s) than FCM, with an overall bias of 2.2-fold (Bland-Altman comparison). Values for artesunate and dihydroartemisinin were most affected. Paradoxical increases in signal at very high concentrations of mefloquine and related compounds were more marked with the DELI assay, suggesting that off-target effects on LDH production may be responsible. Loss of FCM signal due to reinvasion or slow growth was observed in a small number of samples. These results extend previous work on use of flow cytometry to determine antimalarial susceptibility in terms of the number of samples, range of drugs, and comparison with other methods.
author2 Nuffield Department of Clinical Medicine
author_facet Nuffield Department of Clinical Medicine
Charles J. Woodrow
Chirapat Wangsing
Kanlaya Sriprawat
Peter R. Christensen
Francois Nosten
Laurent Rénia
Bruce Russell
Benoît Malleret
format Article
author Charles J. Woodrow
Chirapat Wangsing
Kanlaya Sriprawat
Peter R. Christensen
Francois Nosten
Laurent Rénia
Bruce Russell
Benoît Malleret
author_sort Charles J. Woodrow
title Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions
title_short Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions
title_full Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions
title_fullStr Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions
title_full_unstemmed Comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions
title_sort comparison between flow cytometry, microscopy, and lactate dehydrogenase-based enzyme-linked immunosorbent assay for plasmodium falciparum drug susceptibility testing under field conditions
publishDate 2018
url https://repository.li.mahidol.ac.th/handle/123456789/36314
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