套式PCR检测恶性疟原虫与间日疟原虫方法的建立及其应用研究

Study on the establishment and application of detecting falciparum and plasmodium viviax

目的建立简便、灵敏、低成本的恶性疟原虫与间日疟原虫套式PCR检测方法,并探讨应用于间日疟原虫实验室检测的效果。方法制备抗凝静脉血的干滤纸血滴标本,采用干滤纸5%Chelex-100煮沸法提取疟原虫DNA,并进行套式PCR反应;通过比较PCR检测结果与疟原虫镜检结果,评价套式PCR检测恶性疟原虫与间日疟原虫方法的效果。结果在42份样本中,有34份血样PCR检测结果与镜检结果一致,占81%;5份镜检结果无法判断的血样,PCR检测为阳性或阴性;2份镜检结果为阴性的血样,套式PCR检测为阳性。结论套式PCR检测恶性疟原虫与间日疟原虫结果可靠,比镜检方法灵敏,有望在疟疾的实验室诊断中得到应用。

Objective To establish a simple, convenient, senstive and cheap nested PCR for the detection of Plasmodium falciparum （P. f） and plasmodium viviax （P. v ）, and to apply it for diagnosing plasmodium viviax. Methods Genomic DNA of plasmodium was isolated from filter paper blood samples by 5% chelex-100,and nested PCR was performed. Effect of nested PCR for detecting Plasmodium falciparum and Plasmodium viviax was evaluated by comparing the resulsts between nested PCR and microscopy. Results 42 blood samples were examined, the concordance between nested PCR and microscopy was 81%. 5 samples tmable to be estimated by microscopy,were able to get positive or negative results by nested PCR. In addition, resalts of 2 samples were microscopically negative, showed positive result by nested PCR. Conclusion Results of Plasmodium falciparum and plasmodium by nested PCR were reliable, and it is more sensitive and accurate in detecting Plasmodium species than microscopy.