Ciceron L, Jaureguiberry G, Gay F, Danis M
Service de Parasitologie et Unite INSERM 313, Centre Hospitalier-Universitaire Pitie-Salpetriere, 75013 Paris, France.
J Clin Microbiol. 1999 Jan;37(1):35-8. doi: 10.1128/JCM.37.1.35-38.1999.
We report in this work a highly sensitive and nonradioactive PCR method for the detection of the four species of parasite causing human malaria. Plasmodium-specific primers corresponding to the small-subunit rRNA genes of the malaria parasite were used, and a 291-bp fragment was amplified. Our results showed a high specificity for the four human Plasmodium species, and we were able to detect one parasite in 50 microl of whole blood. The responses of 12 patients infected with Plasmodium falciparum to antimalarial therapy were monitored by PCR diagnosis and examination of thick blood film for at least 20 min by an experienced microscopist. For one patient this study allowed early diagnosis of therapeutic failure, confirmed 7 days later by examination of the thick blood film. A total of 134 samples were examined; 94 were positive by PCR, and among these 68 were positive by thick blood film examination. The sensitivity of the thick blood film was 72.3% compared to PCR and 60.7% compared to dot blot hybridization.
我们在这项工作中报告了一种用于检测导致人类疟疾的四种寄生虫的高灵敏度非放射性PCR方法。使用了与疟原虫小亚基rRNA基因相对应的疟原虫特异性引物，并扩增出了一个291 bp的片段。我们的结果显示对四种人类疟原虫具有高度特异性，并且我们能够在50微升全血中检测到一个寄生虫。通过PCR诊断以及由经验丰富的显微镜检查人员对厚血膜进行至少20分钟的检查，监测了12例感染恶性疟原虫的患者对抗疟治疗的反应。对于一名患者，本研究实现了治疗失败的早期诊断，7天后通过厚血膜检查得到证实。总共检查了134个样本；94个通过PCR呈阳性，其中68个通过厚血膜检查呈阳性。与PCR相比，厚血膜的灵敏度为72.3%，与斑点印迹杂交相比为60.7%。
