A dried blood sample on filter paper is suitable for detecting Plasmodium falciparum gametocytes by reverse transcription polymerase chain reaction

Abstract

The detection of gametocytes in human peripheral blood is one of the most important measures in a malaria survey. We attempted to detect gametocytes of Plasmodium falciparum by reverse transcription polymerase chain reaction (RT-PCR) of dried blood on filter paper. On field samples analysis, the specific RT-PCR products for region 3 of pfg377 mRNA were observed in 67 of 131 falciparum malaria patients. The minimum detection level of RT-PCR-positive samples was 0.03 gametocytes/microl on quantitative real-time RT-PCR. Gametocyte positive rate was not dependent on sex or age. A higher frequency of gametocytes was found in single P. falciparum infection than in mixed species infection (P<0.01). In this study, 47 of the 131 patients were asymptomatic. Eighteen of these 47 patients showed pfg377 mRNA expression. Moreover, four alleles of region 3 of pfg377 were detected in pfg377 mRNA-positive patients and 13 of 67 pfg377 mRNA-positive patients carried more than one gametocyte-producing clone. These results suggest that dried blood on filter paper is a useful for a molecular epidemiologic study of malaria transmission and gametocyte-targeted control.