%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Taylor, B. J.
%A Lanke, K.
%A Banman, S. L.
%A Morlais, Isabelle
%A Morin, M. J.
%A Bousema, T.
%A Rijpma, S. R.
%A Yanow, S. K.
%T A direct from blood reverse transcriptase polymerase chain reaction assay for monitoring Falciparum malaria parasite transmission in elimination settings
%D 2017
%L fdi:010072021
%G ENG
%J American Journal of Tropical Medicine and Hygiene
%@ 0002-9637
%K CAMEROUN
%M ISI:000423197200034
%N 2
%P 533-543
%R 10.4269/ajtmh.17-0039
%U https://www.documentation.ird.fr/hor/fdi:010072021
%> https://horizon.documentation.ird.fr/exl-doc/pleins_textes/divers20-08/010072021.pdf
%V 97
%W Horizon (IRD)
%X We describe a novel one-step reverse transcriptase real-time PCR (direct RT-PCR) for Plasmodium falciparum malaria parasites that amplifies RNA targets directly from blood. We developed the assay to identify gametocyte-specific transcripts in parasites from patient blood samples, as a means of monitoring malaria parasite transmission in field settings. To perform the test, blood is added directly to a master mix in PCR tubes and analyzed by real-time PCR. The limit of detection of the assay on both conventional and portable real-time PCR instruments was 100 parasites/mL for 18S rRNA, and 1,000 parasites/mL for asexual (PFE0065W) and gametocyte (PF14_0367, PFGEXP5) mRNA targets. The usefulness of this assay in field studies was explored in samples from individuals living in a high-transmission region in Cameroon. The sensitivity and specificity of the assay compared with a standard two-step RT-PCR was 100% for 18S rRNA on both conventional and portable instruments. For PF14_0367, the sensitivity and specificity were 85.7% and 70.0%, respectively, on the conventional instrument and 78.6% and 90%, respectively, on the portable instrument. The concordance for assays run on the two instruments was 100% for 18S rRNA, and 79.2% for PF14_0367, with most discrepancies resulting from samples with low transcript levels. The results show asexual and sexual stage RNA targets can be detected directly from blood samples in a simple one-step test on a field-friendly instrument. This assay may be useful for monitoring malaria parasite transmission potential in elimination settings, where sensitive diagnostics are needed to evaluate the progress of malaria eradication initiatives.
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