逆转录套式聚合酶链反应检测庚型肝炎病毒

目的建立敏感、特异的逆转录套式聚合酶链反应（ＲＴ－ｎｅｓｔｅｄ－ＰＣＲ）方法，以检测中国人庚型肝炎病毒（ＨＧＶ）感染。方法根据中国人感染ＨＧＶ者ＮＳ５区部分核苷酸序列分析结果，设计套式ＰＣＲ引物，用于ＨＧＶＲＮＡ的检测，并与用国外报道引物所作的一次ＰＣＲ和套式ＰＣＲ检测结果进行比较。结果共检测标本１３３份。以国外报道引物作一次ＰＣＲ和套式ＰＣＲ检出率分别为８．３％和１１．３％，作者建立的套式ＰＣＲ检出率为１８．０％，对部分ＰＣＲ产物进行序列分析证实为ＨＧＶ特异性基因。结论建立的方法可在中国人群中显著提高ＨＧＶＲＮＡ检出率。

Reverse nested polymerase chain reaction for the detection of hepatitis G virus RNA

Objective To develop a sensitive and specific reverse nested polymerase chain reaction(RT nested PCR)method for the detection of hepatitis G virus (HGV) in Chinese patients. Methods A set of nested PCR primers were designed according to the sequence analysis result of partial NS5 gene of HGV in Chinese patients. Using this set of primers, a RT nested PCR method was developed to detect HGV infection in the Chinese, and its result was compared with one stage PCR and nested PCR, whose primers were designed according to sequences from abroad. Results 133 samples were detected. The positive rate in one stage PCR, nested PCR and our newly developed nested PCR was 8.3%, 11.3% and 18.0% respectively. The specificity of the PCR product was verified by sequence analysis. Conclusion Our developed RT nested PCR method markedly increased the positive rate of HGV RNA among Chinese patients.