Objective To compare and validate Sansure HBV DNA Quantitative Detection Reagent on different detecting platforms.Methods According to the EP series of document issued by the National Committee for Clinical Laboratory Standards (NCCLS) and the Accreditation Criteria for the Quality and Competence of Medical Laboratories,clinical samples were detected by Sansure HBV DNA Quantitative Detection Reagent on Roche LightCycler (R) 480 Ⅱ Real-Time PCR System and ABI 7500 Real-Time PCR System.The non-precision,reportable range,trueness and limit of detection were calculated.Results Roche LightCycler (R)480 Ⅱ Real-Time PCR System:While the concentration level was 2 × 106IU/mL,the CV of within group was 1.38％,and the CV between groups was 0.25％.While the concentration level was 2 × 104IU/mL,the CV of within group was 0.35％,and the CV between groups was 0.79％.The trueness was proved.The biases were calculated by each concentration's,which suggested a good linear correlation in the range of 2.1 × 102IU/mL-2.1 × 107 IU/mL concentration (R2 =0.9994).And the limit of detection was 148 IU/ mL.ABI 7500 Real-Time PCR System:While the concentration level was 2 × 106IU/mL,the CV of within group was 0.70％,and the CV between groups was 1.34％;While the concentration level was 2.5 × 104IU/ mL,the CV of within group was 1.18％,the CV between groups was 3.01％.The trueness was proved.The biases calculated by each concentration'ssuggested a good linear correlation in the range of 2.0 × 102IU/ mL-2.0 × 108IU/mL concentration(R2 =0.9991),and the limit of detection was 203 IU/mL.Conclusion The performance validations of Sansure HBV DNA Quantitative Detection Reagent on both Roche LightCycler (R) 480 Ⅱ Real-Time PCR System and ABI 7500 Real-Time PCR System are consistent with the reagent instruction,and HBV DNA can be detected fast and accurately on both Real-Time PCR Systems.%目的 将圣湘公司乙型肝炎病毒(HBV)核酸定量检测试剂盒(PCR-荧光探针法)在不同荧光定量PCR系统进行性能验证,以给出该试剂盒在不同检测平台上的性能评价.方法 依据美国临床和实验室标准化协会(CLSI)颁布的EP系列文件和《医学实验室质量和能力认可准则》相关文件,应用Roche LightCycler(R)480 Ⅱ实时荧光定量PCR系统及美国ABI 7500型实时荧光定量PCR系统对圣湘公司乙型肝炎病毒(HBV)核酸定量检测试剂盒(PCR-荧光探针法)进行临床样本精密度、正确度、可报告范围和检测下限的性能验证实验.结果 LightCycler(R)480 Ⅱ实时荧光定量PCR系统:2×106 IU/mL浓度水平批内精密度为1.38％,批间精密度为0.25％;2×104IU/mL浓度水平批内精密度为0.35％,批间精密度为0.79％.正确度验证可溯源标准品,偏倚均小于7.5％,符合厂家说明及文件要求.HBV DNA定量结果在2.1×102IU/mL至2.1×107IU/mL浓度水平范围内线性关系良好(R2=0.9994),临床可报告范围为2.1×102IU/mL～2.1×108IU/mL.该系统检测下限为148 IU/mL.ABI 7500型实时荧光定量PCR系统:2×106IU/mL浓度水平批内精密度为0.70％,批间精密度为1.34％.2.5×104IU/mL浓度水平批内精密度为1.18％,批间精密度为3.01％.正确度验证可溯源标准品,偏倚均小于7.5％,符合厂家说明及文件要求.HBV DNA定量结果在2×102IU/mL至2×10sIU/mL浓度水平范围内线性关系良好(R2 =0.9991),临床可报告范围为2×102IU/mL ～2×109IU/mL.该系统检测下限为203 IU/mL.结论 圣湘公司乙型肝炎病毒(HBV)核酸定量检测试剂盒(PCR-荧光探针法)在Roche LightCycler(R)480 Ⅱ实时荧光定量PCR系统及美国ABI 7500型实时荧光定量PCR系统上的性能验证结果均符合试剂说明书性能范围,检测结果无明显差异,均可准确快捷的对HBV DNA定量检测,为后续超敏定量工作奠定基础.
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