M. Menegon et al., Genotyping of Plasmodium falciparum gametocytes by reverse transcriptase polymerase chain reaction, MOL BIOCH P, 111(1), 2000, pp. 153-161
A molecular assay has been developed for the specific detection and genetic
characterisation of Plasmodium falciparum gametocytes in the blood of mala
ria infected individuals. The assay is based on the reverse transcription a
nd polymerase chain reaction (RT-PCR) amplification of the messenger RNA of
gene pfg377, a sexual-stage specific transcript abundantly produced in mat
uring gametocytes. The gene contains four regions of repetitive sequences,
of which region 3 was shown to be the most polymorphic in laboratory clones
and held isolates of the parasite. Analysis of samples of malaria infected
blood by RT-PCR specific for region 3 has enabled identification of multip
le gametocyte-producing clones within single infections. The assay is able
to detect gametocytes below the threshold of microscopic detection, and is
highly specific for its gametocyte targets also in the presence of a vast e
xcess of asexual forms. (C) 2000 Elsevier Science B.V. All rights reserved.