A dried blood sample on filter paper is suitable for detecting Plasmodium falciparum gametocytes by reverse transcription polymerase chain reaction.
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.
Maeno
Yoshimasa
Y
Department of Virology and Parasitology, Fujita Health University School of Medicine, 1-98 Kutsukake, Toyoake, Aichi, Japan. ymaeno@fujita-hu.ac.jp
Nakazawa
Shusuke
S
Dao
Le Duc
le D
Yamamoto
Naoki
N
Giang
Nguyen Duc
ND
Van Hanh
Truong
T
Thuan
Le Khanh
le K
Taniguchi
Koki
K
D000293:Adolescent; D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000818:Animals; D001800:Blood Specimen Collection / Q000295:instrumentation / Q000379:methods*; D002648:Child; D002675:Child, Preschool; D005260:Female; D005374:Filtration / Q000295:instrumentation; D006801:Humans; D007223:Infant; D016778:Malaria, Falciparum / Q000175:diagnosis / Q000453:epidemiology / Q000469:parasitology; D008297:Male; D008875:Middle Aged; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D010209:Paper; D010963:Plasmodium falciparum / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D015800:Protozoan Proteins; D012333:RNA, Messenger