摘要
目的对献血者罗氏COBAS S201血液筛查核酸检测混检系统关键性能指标参数进行验证,为血液病毒核酸检测混检系统性能的确认提供支持数据,并建立科学可行验证方案。方法根据《分子诊断检验程序性能验证指南》(CNAS-GL039:2019)的验证基本要求,对ROCHE COBAS S201核酸检测混检系统的95%检出限、符合率、抗干扰能力、操作人员比对等4个关键参数指标进行全面性能验证,与厂家提供的说明参数进行比对,评估实验室检测系统的实际性能,保证血液检测质量。结果对实验室2台COBAS S201核酸检测混检系统进行厂家声明的95%检出限定值进行单检(PP1)重复检测5次检测,均为100%检出靶核酸,测定下限验证通过。使用单检模式检测5个不同HBV DNA浓度(25~400)IU/mL、5个不同HCV RNA浓度(25~400)IU/mL、5个不同HIV RNA浓度(100~1 000) IU/mL、5个阴性管,结果20支管所有检测值与真实值100%一致,性能参数"符合率"验证通过。对含有脂肪血、溶血、ALT>50 mL/L及梅毒阳性内源性干扰物质的3倍95%检出限定值标本进行重复检测3次,反应率均为100%(3/3),检出率达到100%;对照组(不含或在正常范围干扰物质)在3倍95%检出限这种极低的核酸定值标本同样检出反应性结果,两者的检出率统计学没有显著差异(P>0.05),充分说明献血者血液含有验证浓度的干扰物质(脂肪血、溶血、ALT≥50及梅毒阳性),对检出HBV/HCV/HIV病毒靶核酸无显著影响。同样的仪器同样的试剂同样的标本,由不同的操作人员来进行上样检测,结果出现100%一致性,可初步评估操作人员之间的操作在本次验证并没有差异。结论本研究建立的核酸混检系统的方法学性能指标(95%检出限、符合率、抗干扰能力、操作人员比对)验证方案,具有简易、灵活、科学、可行的特点,可为国内采供血机构同行核酸验证工作提供借鉴与数据支持。
Objective To verify the key performance parameters of Roche Cobas S201 multiplex NAT for blood donors, so as to provide data for the conformation of multiplex NAT performance and establish a scientific and feasible verification scheme. Methods Comprehensive performance verification on four key parameters of ROCHE COBAS S201 multiplex NAT system,including the 95% detection limit, compliance rate, anti-interference ability and operator comparison were conducted. The data were compared with the specifications provided by the manufacturer to evaluate the performance of laboratory testing system and ensure the quality of blood testing. Results 2 sets of COBAS S201 multiplex NAT system in our laboratory were used to perform 5 times of the 95% detection limit value declared by the manufacturer for individual tests. The experimental results showed that the target NAT can be detected 100% and the lower limit of determination was verified. Single-virus NAT was performed to detect 5 different HBV DNA concentrations(25 ~400 IU/mL), 5 different HCV RNA concentrations(25 ~400 IU/mL), 5 different HIV RNA Concentration(100~1 000 IU/mL), and 5 negative tubes. The experimental results showed that all detected values of 20 tubes were 100% consistent with the true value, and the performance parameter "coincidence rate" was verified. Samples containing lipoemia, hemolysis, with ALT>50 mL/L and syphilis-positive endogenous interfering substances with 3 times of the 95% detection limit value were tested for 3 times, and results showed both response rate(3/3) and yielding rate reached 100%. In the control group(with none or in the normal range of interfering substances), reactivity was detected in samples with extremely low concentration of 3 times of the 95% detection limit, showing no significant difference(P>0.05). Above results fully showed that the existence of interference substances(lipidemia, hemolysis, ALT ≥ 50 and syphilis positive) in the blood of donors had no significant effect on the yielding of HBV/HCV/HIV virus target nucleic acids. Different operators performed the sample loading tests with same instrument, reagent and specimen showed 100% consistency in the results. It could be preliminarily assessed that there was no difference in the operations between the operators in this verification. Conclusion The verification scheme of the multiplex NAT system for methodology performance index(95% detection limit, coincidence rate, anti-interference ability, and operator comparison) established in this study, showed simple, flexible, scientific and feasible characteristics and could provide reference and data support for domestic blood transfusion services in acid detection.
作者
黄华霖
郑欣
曾劲峰
HUANG Hualin;ZHENG Xin;ZENG Jingfeng(Shenzhen Longgung Blood Center Station,Shenzhen 518172,China;Shenzhen Blood Center)
出处
《中国输血杂志》
CAS
2021年第2期128-131,共4页
Chinese Journal of Blood Transfusion
关键词
献血者
核酸检测/血液筛查
性能验证
voluntary blood donor
nucleic acid testing/blood screening
performance verification
