PCR检测恶性疟原虫感染蚊方法的建立

Development of a PCR-based method for detection of Plasmodium falciparum infected mosquitoes

目的 建立一种简便的可应用于现场蚊感染恶性疟原虫的PCR检测方法。方法 采用针对恶性疟原虫小亚单位核糖体DNA（SSUrDNA）的特异引物，利用PCR技术，从实验室感染及现场采集的感染恶性疟原虫的蚊基因组DNA中，扩增恶性疟原虫SSUrDNA片段，进行恶性疟原虫的检测。结果 扩增产物经琼脂糖凝胶电泳分析，可检测出特异的约188bp大小的DNA片段，其基因序列与预计序列完全一致。估测每份蚊DNA样本中含有10个以上子孢子即可获得此片段，而对间日疟原虫及未感染蚊DNA不能扩增出此片段。结论 该PCR检测方法可应用于现场恶性疟原虫感染蚊的检测。

Objective To develop a PCR-based method for detection of Plasmodiumfalciparum infected mosquitoes. Methods One pair of primers specific to small subunit ribosomal DNA of P. faciparum were used to amplify the specific SSUrDNA 188 bp fragment of P. faciparum for detecting P. fnciparum infected mosquitoes with PCR-based method. Results As few as 10 sporozoites in one mosquito no matter collected in the field or infected in the lab could be detected with the PCR-based method. In contrast, no such specific 188 bp DNA band was detected in P. vivax and uninfected mosquito. Conclusion The PCR-based method can be used to detect P. faciparum infected mosquitoes collected in the field.