一种简单的检测甲型肝炎病人血清循环免疫复合物中HAV RNA的免疫捕获聚合酶链反应方法

A SIMPLE IMMUNOCAPTURE PCR FOR DETECTION OF HAV RNA IN CIRCULATING IMMUNE COMPLEXES FROM SERUM OF PATIENTS WITH HEPATITIS A

目的 建立一种简单的检测甲型肝炎病人血清循环免疫复合物中HAV RNA的方法。方法 应用差速离心法分离甲肝病人血清中的循环免疫复合物,置于已用抗人μ链包被的离心管中吸咐,再用RT/PCR法扩增,用PAGE银染法检定扩增结果。结果 检测24份甲肝病人血清标本,其中19份为HAV RNA阳性,而检测正常人与乙型肝炎及丙型肝炎病人血清标本均为阴性。结论 该方法简便,敏感性及特异性较高,是检测甲型肝炎病人血清循环免疫复合物中HAV RNA的较好方法之一。

Objective To set up a simple method for detecting HAV RNA in the circulating immune complexes from serum of patient with hepatitis A. Method We adopted differential centrifugation to isolate the circulating immune complexes from serum of patient with hepatitis A. The complexes were added into the micro-centrifuge tube precoated with anti-μ and then RT/PCR was carried out. The amplified products were electrophoresed through PAGE and visualized by silver staining. Results Of 24 serum specimens examined by this method, 19 were HAV RNA positive. But the serum specimens collected from the healthy persons and from the patients with hepatitis B or hepatitis Cexamined by this method, were all negative. Conclusion The method is simple, and its specificity and sensitivity is high. It is one of the method for detecting HAV RNA in the circulating immune complexes from serum of patient with hepatitis A.