HGVRNA逆转录套式聚合酶链反应法的建立及应用  

DEVELOPMENT AND APPLICATION OF A REVERSE TRANSCRIPTION NESTED POLYMERASE CHAIN REACTION FOR DETECTION OF HGV RNA

作　　者：凌斌华[1] 庄辉[1] 李奎[1] 范金水[1] 崔怡辉 汪兴太[2] 杨永芳[3] 

机构地区：[1]北京医科大学微生物学系,北京100083 [2]中国药品生物制品检定所 [3]云南省卫生防疫站

出　　处：《中华肝脏病杂志》1998年第1期21-22,共2页Chinese Journal of Hepatology

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 RNART-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。Objective To establish a reverse transcription nested polymerase chain reaction (RT- nPCR) for the detection of HGV RNA in sera of different populations and patients with various liver diseases.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 this RT- nPCR were 10^(-4),10^(-9),and 10^(-9),respectively.50 HGV RNA negative sera detected by the other RT-nPCR were also negative by this RT-nPCR.The sequence of the amplified RT-nPCR products was identical with that of the target fragment.Of the 97 anti-HGV positive sera tested,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 be used for the detection of HGV RNA in sera of different populations and patients with various liver diseases.

关 键 词：聚合酶链反应 庚型肝炎 核糖核酸 HGV 

分 类 号：R512.63[医药卫生—内科学]