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Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages

Gadalla, Amal A. H. ORCID:, Siciliano, Giualia, Farid, Ryan, Alano, Pietro, Ranford-Cartwright, Lisa, McCarthy, James S., Thompson, Joanne and Babiker, Hamza 2021. Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages. Scientific Reports 11 , 19118. 10.1038/s41598-021-97456-4

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The use of quantitative qRT-PCR assays for detection and quantification of late gametocyte stages has revealed the high transmission capacity of the human malaria parasite, Plasmodium falciparum. To understand how the parasite adjusts its transmission in response to in-host environmental conditions including antimalarials requires simultaneous quantification of early and late gametocytes. Here, we describe qRT-PCR assays that specifically detect and quantify early-stage P. falciparum gametocytes. The assays are based on expression of known early and late gametocyte genes and were developed using purified stage II and stage V gametocytes and tested in natural and controlled human infections. Genes pfpeg4 and pfg27 are specifically expressed at significant levels in early gametocytes with a limit of quantification of 190 and 390 gametocytes/mL, respectively. In infected volunteers, transcripts of pfpeg4 and pfg27 were detected shortly after the onset of blood stage infection. In natural infections, both early (pfpeg4/pfg27) and late gametocyte transcripts (pfs25) were detected in 71.2% of individuals, only early gametocyte transcripts in 12.6%, and only late gametocyte transcripts in 15.2%. The pfpeg4/pfg27 qRT-PCR assays are sensitive and specific for quantification of circulating sexually committed ring stages/early gametocytes and can be used to increase our understanding of epidemiological processes that modulate P. falciparum transmission.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Additional Information: This article is licensed under a Creative Commons Attribution 4.0 International License
Publisher: Nature Research
ISSN: 2045-2322
Date of First Compliant Deposit: 20 September 2021
Date of Acceptance: 25 August 2021
Last Modified: 08 May 2023 22:39

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