PCR检测Pf60.1基因诊断恶性疟实验方法研究

DIAGNOSIS OF FALCIPARUM MALARIA THROUGH DETECTION OF Pf60.1 GENE BY PCR

为探讨通过 PCR技术检测 Pf60 .1基因来诊断恶性疟的可行性 ,采用 PCR技术检测了 4年前采集贮存的 2 0例疑似疟疾病人的血样。阳性对照为恶性疟原虫 FCC1 /HN培养株 ;阴性对照为食蟹猴疟原虫、间日疟原虫及非疟区的正常供血员血样。结果为 FCC1 /HN株、1 2例镜检定为恶性疟和 2例混合感染的血样扩增出长度为 3 1 3 bp～ 3 2 0 bp的特定 Pf60 .1基因片段 ;而阴性对照均未产生 DNA带型。此结果说明检测Pf60 k D中的 Pf60 .1基因片段的方法稳定性好 ,特异性强和敏感性高 ,是恶性疟诊断的理想方法之一。

To explore the possibility for diagnosis of falciparum malaria by detecting Pf60.1 gene, blood samples of 20 suspected malaria cases collected 4 years ago were tested by PCR. In the experiments, the FCC1/HN laboratory strain was selected as positive control, and P. cynomolgi, P. vivax, as well as normal blood samples collected from a blood donor living in non-malaria area as negative controls. The results showed that the FCC1/HN strain, the samples of 12 falciparum malaria cases and 2 mixed infection cases with both P. falciparum and P. vivax produced 313bp～320bp special gene fragments. However the negative control samples did not produce the special DNA band, indicating the method of detecting Pf60.1 fragment of Pf60 kD gene family had favorable stability, specificity and sensitivity. The authors suggest that it would be one of the practical methods for diagnosis of falciparum malaria in epidemiological investigation.