TY - JOUR
T1 - Quantification of female and male Plasmodium falciparum gametocytes by reverse transcriptase quantitative PCR
AU - Schneider, Petra
AU - Reece, Sarah E.
AU - Van Schaijk, Ben C.L.
AU - Bousema, Teun
AU - Lanke, Kjerstin H.W.
AU - Meaden, Cora S.J.
AU - Gadalla, Amal
AU - Ranford-Cartwright, Lisa C.
AU - Babiker, Hamza A.
N1 - Funding Information:
Thanks to Aidan O’Donnell, Ahmed Raza, Liz Peat and Dorothy Armstrong for assistance during the experiments, the Ashworth microparasites group for discussion, and to the anonymous reviewer whose comments helped improve and clarify this manuscript. S.E.R.'s lab is supported by National Environment Research Council ( NE/I015329/1 ), Wellcome Trust ( WT082234MA ), Royal Society , BBSRC and the Centre for Immunity, Infection and Evolution . C.S.J.M. was supported by a BBSRC DTG studentship . K.H.W.L. and T.B. are supported by a grant from the Bill& Melinda Gates Foundation ( OPP1034789 ). L.R.C.’s lab is supported by the Wellcome Trust ( WT091791MA , WT078749MA ).
Publisher Copyright:
© 2015 The Authors. Published by Elsevier B.V.
PY - 2015/1/1
Y1 - 2015/1/1
N2 - The transmission of malaria parasites depends on the presence of sexual stages (gametocytes) in the blood, making the ratio and densities of female and male gametocytes important determinants of parasite fitness. This manuscript describes the development of reverse transcriptase quantitative PCR (RT-qPCR) assays to separately quantify mature female and male gametocytes of the human malaria parasite Plasmodium falciparum, and reveals that Pfs25 mRNA is expressed only in female gametocytes. The female (Pfs25) and male (Pfs230p) gametocyte specific RT-qPCR assays have lower detection limits of 0.3 female and 1.8 male gametocytes per microlitre of blood, respectively, making them more sensitive than microscopy. Accurate quantification of the ratio and densities of female and male gametocytes will increase understanding of P. falciparum transmission and improve the evaluation of transmission blocking interventions.
AB - The transmission of malaria parasites depends on the presence of sexual stages (gametocytes) in the blood, making the ratio and densities of female and male gametocytes important determinants of parasite fitness. This manuscript describes the development of reverse transcriptase quantitative PCR (RT-qPCR) assays to separately quantify mature female and male gametocytes of the human malaria parasite Plasmodium falciparum, and reveals that Pfs25 mRNA is expressed only in female gametocytes. The female (Pfs25) and male (Pfs230p) gametocyte specific RT-qPCR assays have lower detection limits of 0.3 female and 1.8 male gametocytes per microlitre of blood, respectively, making them more sensitive than microscopy. Accurate quantification of the ratio and densities of female and male gametocytes will increase understanding of P. falciparum transmission and improve the evaluation of transmission blocking interventions.
KW - Gametocyte sex ratio
KW - Malaria
KW - PCR
KW - Pfs25 Pfs230p
KW - Sex-specific quantitative
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U2 - 10.1016/j.molbiopara.2015.03.006
DO - 10.1016/j.molbiopara.2015.03.006
M3 - Article
C2 - 25827756
AN - SCOPUS:84926454336
SN - 0166-6851
VL - 199
SP - 29
EP - 33
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology
IS - 1-2
ER -