实时荧光定量 PCR 检测 HBV DNA 的线性范围与最低检出限的验证

目的：试用实时荧光定量 PCR 定量检测乙型肝炎病毒核酸（HBV DNA），求出其线性范围与最低检出限，从而对本实验室检测 HBV DNA 的主要性能进行验证。方法参照相关的文件，通过系列稀释高浓度标本得到超过厂家申明线性范围的系列浓度标本，进行线性范围的验证；通过系列稀释低浓度标本得到超过厂家申明最低检出限的系列浓度标本，进行最低检出限的验证。结果 HBV DNA 检测的线性范围为8．58＆#215;102～8．41＆#215;107 IU/mL，最低检出限为4．07＆#215;102 IU/mL。结论实时荧光定量 PCR 检测 HBV DNA 的线性范围与最低检出限达到预期要求，检测方法和验证方案简便、可行。

Verification of the linear range and the minimum detection limit in real-time fluorescence quantitative PCR for HBV DNA

Objective Use real-time fluorescence quantitative polymerase chain reaction（PCR）to determine HBV DNA,then calculate the linear range and the minimum detection limit,which are the main performance indicators in the laboratory verification. Methods According to the related documents,by serial dilution of high concentration samples,samples of serial concentrations were obtained which were out of the linear range mentioned in the instructions,then verifid the new linear range.By serial dilution of low concentration,samples were obtained,the concentrations of which were lower than the minimum detection limit of provide by the manufacturer,then the new minimum detection limit was verified.Results The linear range of HBV DNA detection was 8.58× 102 -8.41×107 IU/mL,and the minimum detection limit of HBV DNA detection was 4.07 ×102 IU/mL.Conclusion The linear range and the minimum detection limit of Real-time fluorescence quantitative PCR assessed reaches the expected requirement,and the method and validation scheme are simple and feasible.