血站核酸检测系统基于T/CSBT的性能验证探讨

Performance verification of domestic nucleic acid testing system in blood stations based on T/CSBT

目的对血站实验室基于PCR-荧光法的HBV DNA、HCV RNA和HIV RNA-1核酸检测系统进行性能验证,以满足T/CSBT,保障核酸检测的质量。方法用2022年国家卫健委临床检验中心的室间质评标本进行符合率验证,用HBV DNA、HCV RNA和HIV RNA-1标准物质进行分析灵敏度、内源性干扰物质、重复性、抗交叉污染能力、稳定性等性能验证。结果20份室间质评标本检测结果的符合率为100%;HBV DNA、HCV RNA和HIV RNA-13个项目的分析灵敏度验证结果检出率100%,符合T/CSBT;3 g/L浓度脂血和4 g/L浓度溶血对HBV DNA、HCV RNA和HIV RNA-1项目的检出率无影响;重复性验证批内和批间变异系数均<5%,且批内变异系数<批间变异系数,符合T/CSBT;抗交叉污染能力验证40份阴性标本检测结果100%为阴性,40份10000 IU/mL的HBV强阳性标本100%为阳性;稳定性验证结果弱阳性标本检出率100%,试剂在冻融1次和冻融5次后检测结果的变异系数均<5%,检测Ct值在试剂冻融1次和冻融5次之间的差异无统计学意义。结论核酸检测系统及配套试剂检测符合率、分析灵敏度、内源性干扰物质、重复性、抗交叉污染能力、稳定性验证结果均符合T/CSBT,可用于血站实验室血筛实验中的核酸检测。

Objective To explore the performance verification of the Shengxiang automatic NAT system for HBV DNA,HCV RNA and HIV RNA⁃1 using PCR⁃fluorescence in the laboratories of blood stations,in order to meet the requirements of T/CSBT and ensure the quality of nucleic acid detection.Methods Samples used in the external quality assessment(EQA)of National Center for Clinical Laboratories of the year 2022 were taken to verify the concordance.The standard materials of HBV DNA,HCV RNA and HIV RNA-1 were used to verify the analytical sensitivity,endogenous interfering substances,repeatability,anti-cross contamination ability and stability.Results The concordance rate of 20 EQA samples was 100%.The analytical sensitivity of HBV DNA,HCV RNA and HIV RNA-1 were all reactive and met T/CSBT.The yielding of HBV DNA,HCV RNA and HIV RNA-1 was affected lttle with lipemia at 3g/L and hemolysis at 4g/L.The coefficients of variation(CV)of intra-assay and inter-assay which met T/CSBT were all less than 5%,and the intra-assay variation coefficient was less than the inter-assay variation coefficient.The test results of 40 negative samples tested for cross contamination resistance were 100%negative,and 40 positive samples of HBV with 10000 IU/mL were 100%positive.The stability verification results showed that the detection rate of weak positive samples was 100%.The coefficient of variation of the test results of the reagent after 1 and 5 freeze-thaw cycles were less than 5%,and the difference between the detection Ct value of reagent underwent once freeze-thaw and five-time freeze-thaw was not statistically significant.Conclusion The analytical sensitivity,endogenous interfering substances,repeatability,anti-cross contamination ability,stability and the compliance rate of domestic Shengxiang Gene automatic NAT system and supporting reagents by PCR-fluorescence method all meet T/CSBT,so it can be used for nucleic acid detection in blood screening in blood station laboratory.