化学发光法检测抗髓过氧化物酶抗体和抗蛋白酶3抗体

Chemiluminescent Immunoassay for the Detection of Anti-myeloperoxidase and Anti-proteinase 3 Autoantibodies

目的研究全自动、定量和随机上样化学发光法(chemiluminescent immunoassay,CLIA)检测抗髓过氧化物酶(myeloperoxidase,MPO)抗体和抗蛋白酶3(proteinase 3,PR3)抗体的临床应用价值,为临床抗MPO抗体和抗PR3抗体检测方法的选择提供参考。方法对临床242例常规申请抗中性粒细胞胞浆抗体(anti-neutrophil cytoplasmic autoantibodies,ANCA)检测的样本采用间接免疫荧光法(immunofluorescent assay,IFA)筛查的同时,应用CLIA和酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)平行检测抗MPO抗体和抗PR3抗体。对上述检测结果进行统计学分析。结果 CLIA与ELISA针对抗MPO抗体的检测表现出良好的一致性和符合率,其中总符合率为93.8%,阳性符合率81.9%,阴性符合率98.8%,一致性结果显示kappa=0.845,受试者工作特征曲线(receiver operating characteristic curve,ROC曲线)以下面积(area under the curve,AUC)为0.966。两种方法学在检测抗PR3抗体时一致性和符合率表现较低,其中总符合率为86.8%,阳性符合率为35.1%,阴性符合率为96.1%。ROC曲线分析显示AUC为0.839;抗PR3抗体两种方法检测结果不一致的样本经第三方试剂验证结果显示,CLIA检测结果与第三方试剂验证结果差异无统计学意义(P>0.05),而ELISA检测结果与第三方试剂验证结果差异存在统计学意义(P<0.05)。CLIA检测结果与IFA检测结果阳性符合率为60.8%、阴性符合率为94.8%、总符合率为79.8%。而ELISA检测结果与IFA检测结果阳性符合率、阴性符合率和总符合率则分别为73.6%、77.2%和75.6%。结论与ELISA比较,CLIA检测抗MPO抗体具有良好的一致性和符合率,而检测抗PR3抗体时一致性和符合率偏低。在检测抗PR3抗体和抗MPO抗体时,CLIA的特异性均优于ELISA。建议实验室开展抗MPO抗体和抗PR3抗体检测方法选择时,应同时结合IFA抗体筛查试验、样本临床信息以及方法学自身特点等因素进行严格的判断和评价。

Objective To evaluate the clinical performance of an automated, quantitative and random-ac- cessed Chemiluminescent Immunoassay （CLIA） for testing anti-MPO and anti-PR3 autoantibodies.Methods A total of 242 clinical serum samples were screened by immunofluorescence assay （IFA） for anti-neutrophil autoantibody （ANCA）and then tested with CLIA and enzyme-linked immunosorbent assay （ELISA） for anti-MPO and anti-PR3 autoantibodies in parallel. Results CLIA and ELISA showed ex- cellent agreement for anti-MPO. The overall agreement, negative agreement, and positive agreement were 93.8%, 98.8%, and 81.9% respectively. Receiver operating characteristics （ROC） analysis showed that area under the curve （AUC） was 0. 966 for anti-MPO. While a relatively poor agreement between CLIA and ELISA for anti-PR3 was found. The overall agreement, negative agreement, and positive agreement were 86. 8%, 96. 1%, and 35. 1%. Receiver operating characteristics （ROC） analysis showed that the AUC was 0. 839 for anti-PR3. All CLIA and ELISA discrepant samples for anti-PR3 were retested with the reagents from other manufacturer （the validation reagent）. No significant difference was observed be- tween CLIA and the validation reagent. But a significant difference was detected in the test results between ELISA and the validation reagents. The positive, negative and total agreement between CLIA and IFA were 60. 8%, 94. 8%, and 79. 8%, while the positive, negative, and total agreement between ELISA and IFA were 73.6%, 77.2% and 75.6%. Conclusions Excellent agreement can be found for anti-MPO but a relatively poor agreement for anti-PR3 between CLIA and ELISA has been found. The specificity of CLIA is superior to ELISA for both anti-MPO and anti-PR3. We recommend that careful verification and validation should be performed based on IFA screening test, clinical diagnosis and assay features before choosin~ a test method for anti-MPO and anti-PR3.