Maeno Yoshimasa, Nakazawa Shusuke, Dao Le Duc, Yamamoto Naoki, Giang Nguyen Duc, Van Hanh Truong, Thuan Le Khanh, Taniguchi Koki
Department of Virology and Parasitology, Fujita Health University School of Medicine, 1-98 Kutsukake, Toyoake, Aichi, Japan.
Acta Trop. 2008 Aug;107(2):121-7. doi: 10.1016/j.actatropica.2008.05.001. Epub 2008 May 8.
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.
检测人体外周血中的配子体是疟疾调查的最重要措施之一。我们试图通过对滤纸上的干血进行逆转录聚合酶链反应（RT-PCR）来检测恶性疟原虫的配子体。在现场样本分析中，131例恶性疟患者中有67例观察到了pfg377 mRNA第3区的特异性RT-PCR产物。定量实时RT-PCR检测RT-PCR阳性样本的最低检测水平为0.03个配子体/微升。配子体阳性率与性别或年龄无关。单一恶性疟原虫感染中配子体的出现频率高于混合感染（P<0.01）。在本研究中，131例患者中有47例无症状。这47例患者中有18例显示出pfg377 mRNA表达。此外，在pfg377 mRNA阳性患者中检测到了pfg377第3区的4个等位基因，67例pfg377 mRNA阳性患者中有13例携带不止一个产生配子体的克隆。这些结果表明，滤纸上的干血对于疟疾传播的分子流行病学研究和以配子体为靶点的控制是有用的。
