应用分子信标荧光PCR定量检测乙型肝炎病毒核酸的评价

Evaluation of a Molecular Beacon Fluorescence PCR Kit for Quantitative Detection of Hepatitis B Virus Nucleic Acid

目的:分子信标(Molecular Beacon)是近年新发展的一种荧光标记发夹形寡核苷酸探针。本文目的在于对国产分子信标荧光PCR定量检测HBV核酸试剂盒(厦门泰伦生物工程有限公司的“乙型肝炎病毒荧光PCR定量检测试剂盒”)的主要性能进行评价。方法:以Roche公司的乙型肝炎病毒(HBV)定量检测试剂盒AMPLICOR HBV MONITOR^(TM) Test Kit为参比,对分子信标定量检测临床血清标本HBV DNA的结果,进行比较和分析。结果:根据两种试剂盒定量检测34份乙型肝炎和非肝炎血清及一份系列10倍稀释血清(含高水平HBV DNA)的数据,并以Roche试剂盒的结果为参比标准,分子信标试剂盒定量检测HBV DNA的相对敏感性、特异性和符合率均为100％(分别为:19/19,15/15和34/34);其定量检测HBV DNA的线性范围和下限分别为10~3～10~8 copies/ml和10~3 copies/ml;两种试剂盒定量检测HBV DNA的相关系数(r)=0.987。结论:初步结果表明:该国产分子信标荧光PCR定量检测HBV核酸试剂盒的主要性能,与Roche公司的AM-PLICOR HBV MONITOR^(TM) Test试剂盒相似,并具有较后者更宽的线性定量范围。

Objective: To evaluate a molecular beacon fluorescence PCR kit for quantitative detection of hepatitis B virus nucle-ic acid (produced by Xiamen Tailun Biothec Co, Fujian, China) .Methods: This molecular beacon kit was compared with Roche'AM-PIJCOR HBV MONITOR?Test' kit for quantitative detection of HBV DNA in clinical serum specimens.Results: Based on the data that the HBV DNA levels in 34 sera from patients with and without hepatitis B and a serial 10 fold diluted serum (containing HBV DNA) detected by the two kits and using the results by the Roche kit as reference Standard, the relative specificity, sensitivity and over-all accordance of the molecular beacon kit were all 100% (19/19, 15/15, and 34/34, respectively); the linear quantitative range and real detection lower limit of this kit were itf ~ itf and itf HBV DNA copies/rol, respectively; the correlation coefficient (r) of HBV DNA quantitative detection between the two kits was 0.987.Conclusion: Preliminary results indicate that the main efficacy of the molecular beacon fluorescence PCR kit for guantitative detection of hepatitis B virus nucleic acid is similar to the Roche' AM-PLICOR HBV MONTTOR?Test' kit for the same purpose and the former kit has a wider h'near quantitative range.