核酸检测系统的性能验证

Performance verification of four nucleic acid detection systems:A comparitve study

目的对本实验室在用的4套核酸检测系统Tigris、Panther、ChiTaS BSS1200和cobas S201的分析性能进行验证,保证其性能满足血液筛查的要求,并对4套系统的结果一致性进行比较。方法参照ISO15189《医学实验室质量和能力认可准则在分子诊断领域的应用说明(CNAS-CL02-A009:2018)》、《分子诊断检验程序性能验证指南(CNAS-GL039:2019)》以及《血站技术操作规程(2019)版》相关文件的要求,核酸检测项目需要验证的参数包括:分析灵敏度验证、设备间性能比对验证、抗干扰能力、抗交叉污染能力。结果Tigris检测系统HBV DNA、HIV RNA、HCV RNA的95%检出限分别为:2.013 IU/mL、13.039 IU/mL、2.278 IU/mL;Panther检测系统HBV DNA、HIV RNA、HCV RNA 95%检出限分别为:4 IU/mL、10.21 IU/mL、2.077 IU/mL;ChiTaS BSS1200检测系统HBV DNA、HIV RNA、HCV RNA的95%检出限分别为:2.995 IU/mL、30.952 IU/mL、12.008 IU/mL;Cobas S201检测系统HBV DNA、HIV RNA、HCV RNA 95%检出限分别为:0.99 IU/mL、32.24 IU/mL、3.39 IU/mL;在同一厂家相同检测系统2套设备间Tigris和Panther的性能比对验证中,2套Tigris检测系统结果与性能验证血清盘完全一致,符合率100%,Kappa值为1,无交叉污染发生;1号Panther检测系统结果符合率为100%,2号Panther检测系统结果符合率为98%,Kappa值为0.961,无交叉污染发生。低、中度溶血标本、脂肪血标本对低浓度标本检出无影响。4套系统比对后符合率为100%,灵敏度为100%、特异性为100%。结论4套检测系统都是有着极高灵敏度的全自动核酸检测系统,相同检测系统不同设备间检测性能无显著性差异,检测系统的分析灵敏度满足厂商说明书给出的检出限,可满足检测要求。Panther检测系统HCV RNA、HIV RNA分析灵敏度高于Tigris,HBV DNA分析灵敏度低于Tigris,符合试剂说明书要求。溶血、乳糜血对检测结果无显著影响。

Objective To compare the analytical performance of Tigris,Panther,ChiTaS BSS1200 and cobas S201 system to see if they satisfy the requirements of blood screening and to know the concordance of the results presented by these four systems.Methods According to the relevant documents of ISO15189 and Standard Operating Procedure of Blood Station(2019),the parameters needed to be verified for nucleic acid tests(NAT)included:analytical sensitivity verification,system compare test,anti-jamming capability and anti-cross-contamination ability.Results The 95%detection limits of Tigris,Panther,ChiTaS BSS1200 and Cobas S201 for HBV-DNA(IU/mL),HIV-RNA(IU/mL)and HCV-RNA(IU/mL)were 2.013 vs 4 vs 2.995 vs 0.99,13.039 vs 10.21 vs 30.952 vs 32.24,and 2.278 vs 2.077 vs 12.008 vs 3.39,respectively.In the performance comparison verification between NAT systems,the results of the two sets of Tigris systems were in full accordance with serum plate,with a concordance rate of 100%,Kappa value of 1,and none cross-contamination.The concordance rate of No.1 Panther system was 100%,and No.298%,with Kappa value of 0.961 and none cross-contamination.Hemolytic samples(5 g/L Hb concentration)and lipemic blood samples(13.81 mmol/L TG concentration)had no significant effect on the detection of low-concentration samples.Conclusion No significant differences in the performance of NAT systems were notable by devices,as the four systems were fully automated with high sensitivity,which can fully satisfy the blood screening requirements.Panther system demonstrates superior analysis sensitivity in HCV-RNA/HIV-RNA and lower in HBV DNA,but also in required criteria,as compared to Tigris system.Neither hemolysis nor lipemic blood had any significant effect on the test results.