Plasmodium falciparum: expression of gametocyte-specific genes in monolayer cultures and malaria-positive blood samples.
In order to facilitate molecular epidemiological studies on the transmission of the malaria parasite Plasmodium falciparum a sensitive assay for gametocyte detection based on RT-PCR was developed. The transcription of the sexual stage-specific genes Pfs16, Pfs48/45, Pfs230, and Pfs25, as well as the sexual stage- and sporozoite-specific S 18S rRNA, was detected by RT-PCR. S 18S rRNA was present in seven of nine P. falciparum-positive blood samples, despite the lack of microscopic detection of gametocytes and a parasitemia below 0.1%. Expression of the other four gametocyte-specific genes was detected less frequently in malaria-positive blood samples. These findings indicate that RT-PCR of S 18S rRNA is a highly sensitive method for gametocyte detection and, furthermore, that gametocytes are present in the peripheral blood of most malaria carriers, even if the parasitemia is below 0.1%. To determinate the expression pattern of sexual stage-specific genes in more detail, RT-PCR was performed at consecutive time points of highly synchronized monolayer cell cultures. Transcripts of all examined genes except Pfs25 were detected directly after invasion of merozoites of the strain NF54 in red blood cells.
Niederwieser
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Swiss Tropical Institute, Basel, Switzerland.
Felger
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Beck
H P
HP
D000818:Animals; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000302:isolation & purification; D004912:Erythrocytes / Q000469:parasitology*; D005786:Gene Expression Regulation; D006801:Humans; D016778:Malaria, Falciparum / Q000097:blood / Q000175:diagnosis / Q000469:parasitology*; D018512:Parasitemia / Q000097:blood / Q000175:diagnosis / Q000469:parasitology*; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D016053:RNA, Protozoan / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D012337:RNA, Ribosomal, 18S / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D018076:DNA, Complementary; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S