摘要
目的 建立鉴别人体四种疟原虫的套式 PCR技术 ,检测我国西南部分疟区疟原虫的感染情况。方法 选用两组疟原虫引物 ,扩增 SSU r DNA特定片段 ,经琼脂糖凝胶电泳、溴化乙锭染色 ,紫外检测凝胶分析仪观察并摄像。结果 采用上述方法扩增出间日疟、恶性疟和三日疟分别为 10 4bp、10 2 bp和 115 bp特异片段。所检测的 138份疟疾患者血样中 ,四川名山县 95份均为间日疟单纯感染 ,筠连县 37份中 10份为与间日疟和 /或恶性疟呈混合感染的三日疟 ,云南金平县的 6份均为合并恶性疟的两种或两种以上疟原虫混合感染。结论 所建立的套式PCR检测系统具有敏感性高、特异性好和稳定可靠等优点 ,在疟疾诊断、虫种鉴别、混合感染的判定、大规模流行病学研究和疫情监控预测等方面具有实际应用价值。
Objective To establish a method for identification of four species of human malaria parasites by nested PCR in the southwest part of China. Methods Two pair genus primers were chosen to amplify SSUrDNA fragment from malarial blood samples PCR products were electrophoresed on 2% agarose gels, stained with EB to confirm amplification. Results A total of 138 malarial blood samples were examined by nested PCR, About 103bp, 101bp and 115bp DNA fragments expected were amplified from the blood samples with P.vivax, P.falciparum, and P.malariae and P.ovale as positive controls. It was found that all 95 of the samples from Mingshan County of Sichuan Province showed existence P.vivax,that 10 of 37 samples from Junlian County of Sichuan showed P.malariae mixed with P.vivax and /or P.falciparum, and that all 6 of the samples from Jinping County of Yunnan Province showed P.falciparum mixed with two or more species. Conclusion The results demonstrate that this system is sensitive, specific, reliable, and had the advantage of detecting four mixed infections of malaria at the same time, thus providing a practical tool in the diagnosis and epidemiological survey of malaria.
出处
《华西医科大学学报》
CAS
CSCD
北大核心
2002年第3期452-455,共4页
Journal of West China University of Medical Sciences
基金
日本丰田基金资助
关键词
间日疟
恶性疟
三日疟
套式PCR
Plasmodium vivax Plasmodium falciparum Plasmodium malariae Nested polymerase chain reaction
