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

Abstract 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 co...

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Autores principales:	Amal A. H. Gadalla, Giulia Siciliano, Ryan Farid, Pietro Alano, Lisa Ranford-Cartwright, James S. McCarthy, Joanne Thompson, Hamza A Babiker
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e868c3a6213e4d65811a67f617760857
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spelling	oai:doaj.org-article:e868c3a6213e4d65811a67f6177608572021-12-02T18:51:28ZReal-time PCR assays for detection and quantification of early P. falciparum gametocyte stages10.1038/s41598-021-97456-42045-2322https://doaj.org/article/e868c3a6213e4d65811a67f6177608572021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97456-4https://doaj.org/toc/2045-2322Abstract 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.Amal A. H. GadallaGiulia SicilianoRyan FaridPietro AlanoLisa Ranford-CartwrightJames S. McCarthyJoanne ThompsonHamza A BabikerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Amal A. H. Gadalla Giulia Siciliano Ryan Farid Pietro Alano Lisa Ranford-Cartwright James S. McCarthy Joanne Thompson Hamza A Babiker Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages
description	Abstract 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.
format	article
author	Amal A. H. Gadalla Giulia Siciliano Ryan Farid Pietro Alano Lisa Ranford-Cartwright James S. McCarthy Joanne Thompson Hamza A Babiker
author_facet	Amal A. H. Gadalla Giulia Siciliano Ryan Farid Pietro Alano Lisa Ranford-Cartwright James S. McCarthy Joanne Thompson Hamza A Babiker
author_sort	Amal A. H. Gadalla
title	Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages
title_short	Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages
title_full	Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages
title_fullStr	Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages
title_full_unstemmed	Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages
title_sort	real-time pcr assays for detection and quantification of early p. falciparum gametocyte stages
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/e868c3a6213e4d65811a67f617760857
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Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages

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