应用WS/T 514-2017新标准建立并验证输血前ELISA定性筛查八项的检出限

Establishing and Verifing Detection Limit for the Eight Items before Blood Transfusion with Enzyme-Linked Immunosorbent Assay by the WS/T 514-2017 Industry Standard Documents

目的根据医院等级评定及ISO15189:2012的要求,对输血前酶联免疫法定性筛查八项的检出限进行评价。方法根据2017年新出台的卫生行业标准WS/T 514-2017∶《临床检验方法检出能力的确立和验证》中的概率单位方案建立输血前八项项目的检出限(limit of detection,LoD):在同一套检测系统中,使用2个试剂批号,每个批号连续4天,每天对5个评估浓度值标本重复检测5次,计算对应的命中率,将命中率转化为概率单位,与对应浓度值制作回归模型,将命中率为95%对应的概率单位1.645代入方程式中,所得的浓度值即为LoD估计值;对上述检出限估计值进行验证,连续3天检测对2个LoD声明浓度附近的标本(由标准物质稀释所得)重复检测4次,计算≥LoD声明的阳性结果百分比,≥临界值87%,则验证成功。结果各个项目的 LoD估计值为:乙型肝炎表面抗原0.100IU/ml,乙型肝炎表面抗体9.642 mIU/ml,乙型肝炎e抗原0.666NCU/ml,乙型肝炎e抗体3.700NCU/ml,乙型肝炎核心抗体0.786IU/ml,丙型肝炎病毒抗体0.506NCU/ml,梅毒螺旋体特异性抗体2.236mIU/ml,人类免疫缺陷病毒0.135NCU/ml。检出限估计值验证通过。结论验证项目在该实验室的检测方法及检测系统中的检出限满足要求。

Objective To evaluate the limit of detection of eight enzyme-linked immunosorbent assay（ELISA）according to hospital grade assessment and ISO15189：2012.Methods According to the new health industry standard WS/T 514-2017：＂Establishment and verification of detection capability for clinical laboratory measurement procedures＂,the limit of detection（LoD）was established,in the sameset of detection system,using two reagent lot,each lot for 5 consecutive days 4 consecutive days to assess the value of the concentration of five specimens were detected repeatedly,calculated the corresponding hit rate,then transform into probability units,and the corresponding concentration value production regression model,the hit rate of 95% corresponds to the probability unit 1.645 substituted into the equation,the resulting concentration value was LoD estimates.The detection limit values were tested for 3 consecutive days of detection of two LoD concentrations near the declared concentration of the sample（diluted by the standard material）was detected 4 times repeatedly to calculate the positive result was greater than or equal to the percentage of LoD statement,greater than or equal to the critical value of 87%,then verified success.Results HBsAg：0.100 IU/ml,HBsAb：9.642 mIU/ml,HBeAg：0.666 NCU/ml,HBeAb：3.700 NCU/ml,HBcAb：0.786 IU/ml,HCV：0.506 NCU/ml,TP：2.236 mIU/ml and HIV：0.135 NCU/ml.The detection limit estimates were passed.Conclusion The verification limit of the verification project in the testing method and detection system of the laboratory meet the requirements Objective.