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dc.contributor.authorChristensen, Peteren_US
dc.contributor.authorBozdech, Zbyneken_US
dc.contributor.authorWatthanaworawit, Wanitdaen_US
dc.contributor.authorImwong, Mallikaen_US
dc.contributor.authorRénia, Laurenten_US
dc.contributor.authorMalleret, Benoîten_US
dc.contributor.authorLing, Clareen_US
dc.contributor.authorNosten, Françoisen_US
dc.identifier.citationChristensen, P., Bozdech, Z., Watthanaworawit, W., Imwong, M., Rénia, L., Malleret, B., Ling, C. & Nosten, F. (2022). Reverse transcription PCR to detect low density malaria infections. Wellcome Open Research, 6, 39-.
dc.description.abstractBackground: Targeted malaria elimination strategies require highly sensitive tests to detect low density malaria infections (LDMI). Commonly used methods for malaria diagnosis such as light microscopy and antigen-based rapid diagnostic tests (RDTs) are not sensitive enough for reliable identification of infections with parasitaemia below 200 parasites per milliliter of blood. While targeted malaria elimination efforts on the Thailand-Myanmar border have successfully used high sample volume ultrasensitive quantitative PCR (uPCR) to determine malaria prevalence, the necessity for venous collection and processing of large quantities of patient blood limits the widespread tractability of this method. Methods: Here we evaluated a real-time reverse transcription PCR (RT-qPCR) method that reduces the required sample volume compared to uPCR. To do this, 304 samples collected from an active case detection program in Kayin state, Myanmar were compared using uPCR and RT-qPCR. Results: Plasmodium spp. RT-qPCR confirmed 18 of 21 uPCR Plasmodium falciparum positives, while P. falciparum specific RT-qPCR confirmed 17 of the 21 uPCR P. falciparum positives. Combining both RT-qPCR results increased the sensitivity to 100% and specificity was 95.1%. Conclusion: Malaria detection in areas of low transmission and LDMI can benefit from the increased sensitivity of ribosomal RNA detection by RT-PCR, especially where sample volume is limited. Isolation of high quality RNA also allows for downstream analysis of malaria transcripts.en_US
dc.description.sponsorshipAgency for Science, Technology and Research (A*STAR)en_US
dc.relation.ispartofWellcome Open Researchen_US
dc.rights© 2022 Christensen P et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en_US
dc.subjectScience::Biological sciencesen_US
dc.titleReverse transcription PCR to detect low density malaria infectionsen_US
dc.typeJournal Articleen
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.description.versionPublished versionen_US
dc.subject.keywordsLow Density Malaria Infectionen_US
dc.description.acknowledgementThis study was supported by the Wellcome Trust through a strategic award “Eliminating malaria to counter artemisinin resistance” [101148,]. Funding was also obtained from the following sources; the Bill and Melinda Gates Foundation; The Singapore Immunology Network, A*STAR core fund; the NUHS start-up funding [NUHSRO/2018/006/SU/01]; NUHS seed fund [NUHSRO/2018/094/T1]; the Wellcome Trust Mahidol University Oxford Tropical Medicine Research Programme and the New Zealand HRC eASIA [17/678] project grant.en_US
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