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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\n                    <jats:italic>P. falciparum<\/jats:italic>\n                    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\n                    <jats:italic>pfpeg4<\/jats:italic>\n                    and\n                    <jats:italic>pfg27<\/jats:italic>\n                    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\n                    <jats:italic>pfpeg4<\/jats:italic>\n                    and\n                    <jats:italic>pfg27<\/jats:italic>\n                    were detected shortly after the onset of blood stage infection. In natural infections, both early (\n                    <jats:italic>pfpeg4<\/jats:italic>\n                    \/\n                    <jats:italic>pfg27<\/jats:italic>\n                    ) and late gametocyte transcripts\n                    <jats:italic>(pfs25<\/jats:italic>\n                    ) were detected in 71.2% of individuals, only early gametocyte transcripts in 12.6%, and only late gametocyte transcripts in 15.2%. The\n                    <jats:italic>pfpeg4<\/jats:italic>\n                    \/\n                    <jats:italic>pfg27<\/jats:italic>\n                    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\n                    <jats:italic>P. falciparum<\/jats:italic>\n                    transmission.\n                  <\/jats:p>","DOI":"10.1038\/s41598-021-97456-4","type":"journal-article","created":{"date-parts":[[2021,9,27]],"date-time":"2021-09-27T06:20:00Z","timestamp":1632723600000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Real-time PCR assays for detection and quantification of early P. falciparum gametocyte stages"],"prefix":"10.1038","volume":"11","author":[{"given":"Amal A. 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