套式PCR扩增SSU rDNA特定片段检测云南金平县疟原虫感染的研究

目的　采用套式 PCR系统诊断、鉴别人体疟原虫感染。　方法　采用已建立的套式 PCR系统扩增 SSU r DNA特定片段检测云南金平县恶性疟镜检阳性患者的 6份血样 ,并设阳性与阴性对照。　结果　间日疟原虫、恶性疟原虫和三日疟原虫感染血样中分别扩增出 10 4 bp、10 2 bp和 115 bp预期大小的特定扩增带。正常人血、人源弓形虫、杜氏利什曼原虫 DNA及灭菌双蒸水均未产生特异扩增带。 6份血样中检出 4份 P.v.、P.f.和 P.m.的混合感染 ,1份 P.v.和 P.f .及 1份 P.f .和 P.m.的混合感染。　结论　该系统特异、灵敏、稳定 ,在诊断疟疾的同时可准确地判定混合感染 ,对疟疾的诊断、大规模流行病学研究及疫情监控等具有实际应用价值。

STUDIES ON DIAGNOSIS OF HUMAN MALARIA PARASITES BASED ON AMPLIFYING SPECIFIC SSU rDNA FRAGMENT WITH NESTED PCR IN JINPING, YUNNAN

Objective To diagnose and identify human malaria parasites by nested PCR. Methods To detect human malaria parasites of 6 blood samples obtained from Plasmodium falciparum patients in Jinping, Yunnan by nested PCR system established that targets specific sequence in the SSU rDNA, using blood samples of positive controls and negative controls. Results 104 bp,102 bp and 115 bp DNA fragments expected were amplified from the blood samples of patients with P. vivax, P. falciparum and P. malariae infections respectively. No amplification was observed with DNA of nomal blood, T. gondii, L. donovani and distilled water. 6 cases of mixed Plasmodium infections were found by nested PCR. Conclusion It is suggested that this method is specific, sensitive, stable in detecting mixed infections while diagnose malaria. It provide a powerful tool in diagnosis and epidemiology of malaria.