化学发光免疫分析法检测弓形虫抗体的性能验证

Validation of Chemiluminescence Immunoassay for Detection of Toxoplasma Gondii Antibodies

目的验证化学发光免疫分析法(CLIA)测定弓形虫抗体的性能。方法收集2019年1月至2021年5月南京医科大学附属无锡人民医院收治的15例弓形虫抗体阳性患者及同期体检合格的15例健康人群的血清样本,利用全自动化学发光分析仪及其配套试剂、校准品,获得检测弓形虫抗体的正确度、精密度、临界值以及与其他方法[酶联免疫吸附试验(ELISA)法]的一致性等指标,以评估该方法的性能。结果使用室间质量评价样品检测结果的阴性符合率、阳性符合率均为100%。CLIA检测弓形虫免疫球蛋白(Ig)M批内精密度变异系数(CV)在高低浓度水平分别为2.03%与2.17%,批间精密度CV在高低浓度水平分别为2.36%与2.45%;弓形虫IgG批内精密度CV在高低浓度水平分别为0.88%与4.55%,批间精密度CV在高低浓度水平分别为1.40%与4.34%。重复测定判断值附近±20%的标本20次,符合率100%。CLIA与ELISA法检测弓形虫IgG的Kappa值为0.933,一致百分比为96.67%。结论化学发光免疫法检测弓形虫抗体性能指标达标,具有灵敏度高、特异性强、操作简便、自动化程度高等优势,可应用于临床检验。

Objective To verify the performance of chemiluminescence immunoassay(CLIA)for the determination of Toxoplasma gondii antibody.Methods The serum samples of 15 patients with Toxoplasma gondii antibody positive and 15 healthy people who passed the physical examination in the same period were collected from Wuxi People′s Hospital affiliated to Nanjing Medical University from Jan.2019 to May 2021.The accuracy,precision,critical value and consistency with other methods[enzyme linked immunosorbent assay(ELISA)]of automatic chemiluminescence analyzer and its supporting reagents and calibrators for Toxoplasma gondii antibody were obtained,to evaluate the performance of the method.Results The negative and positive coincidence rates of the test results of the samples by the external quality assessment were both 100%.The intra-batch precision coefficient of variation(CV)of Toxoplasma gondii immunoglobulin(Ig)M was 2.03%and 2.17%at the high and low concentration level respetively,and the inter-batch precision CV was 2.36%and 2.45%at the high and low concentration level,respectively;the intra-batch precision CV of Toxoplasma gondii IgG was 0.88%and 4.55%at high and low concentration levels respectively,and the inter-batch precision CV was 1.40%and 4.34%at high and low concentration level respectively.Determination of±20%samples near the judgment value was done for 20 times,and the coincidence rate was 100%.The Kappa value of Toxoplasma gondii IgG detected by CLIA and ELISA method was 0.933,and the consistent percentage was 96.67%.Conclusion CLIA has the advantages of high sensitivity,strong specificity,simple operation and high degree of automation for the detection of Toxoplasma gondii antibodies,and can be applied in the clinical examination.