关键词:逆转录套式聚合酶链反应 庚型肝炎病毒 庚型肝炎病毒核糖核酸 抗-HGV
【摘要】 目的 建立HGV RNA逆转录套式聚合酶链反应法(RT-nPCR)并应用于我国不同人群HGV感染的检测。 方法 根据中国株HGV 5'端非编码区序列设计引物,建立HGV RNA RT-nPCR法。 结果 检测3份HGV RNA阳性血清,其最终阳性稀释度分别为10-4、10-9和10-9;检测50份HGV RNA阴性血清均为阴性。检测97份抗-HGV阳性血清,其中HGV RNA检出率为65.97%。 结论 应用中国株HGV 5'端非编码区序列设计引物,建立的HGV RNA RT-nPCR法灵敏度高,特异性好,可用于我国各类人群和各型肝炎患者HGV RNA的检测。
DEVELOPMENT ANDAPPLICATION OF A REVERSE TRANSCRIPTION NESTED POLYMERASE CHAIN REACTION FOR DETECTION OFHGV RNA
Ling Binhua, Zhuang Hui, Wang Xintai, et al. Department of Microbiolgy, Beijing Medical University, Beijing 100083
【Abstract】 Objective Toestablish a reverse transcription nested polymerase chain reaction (RT-nPCR) for thedetection of HGV RNA in sera of different populations and patients with various liverdiseases. Methods Two pairs of primers were designed based on the sequence of the 5'non-coding region of a Chinese HGV isolate. Results The final HGV RNA positive dilutions of 3 sera tested by thisRT-nPCR were 10-4, 10-9, and 10-9, respectively. 50 HGV RNA negative sera detected by theother RT-nPCR were also negative by this RT-nPCR. The sequence of the amplified RT-nPCRproducts was identical with that of the target fragment. Of the 97 anti-HGV positive seratested, 64 (65.97%) were HGV RNA positive. Conclusion The HGV RNA RT-nPCR using the primers derived from the 5'non-coding region of a Chinese HGV isolate has high sensitivity and specificity. It can beused for the detection of HGV RNA in sera of different populations and patients withvarious liver diseases.
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