纳米磁微粒化学发光免疫分析法检测过敏原特异性免疫球蛋白E性能评估

Performance of magnetic nanoparticle chemiluminescence immunoassay in detection of allergen-specific immunoglobulin E

目的评估纳米磁微粒化学发光免疫分析法(NM-CLIA)检测屋尘螨(国际过敏原代码D1)和粉尘螨(国际过敏原代码D2)过敏原特异性免疫球蛋白E(s Ig E)抗体的相关性能。方法分别运用NM-CLIA和免疫荧光法检测489例在上海交通大学医学院附属苏州九龙医院就诊的疑似过敏患者的血清样本,其中D1疑似过敏244例、D2疑似过敏245例,采用χ~2检验和Kappa检验评价2种方法检测D1和D2 s Ig E抗体的相关性。按照美国临床实验室标准化协会标准方法验证NM-CLIA检测D1和D2 s Ig E抗体的最低检出限(Lo D)、线性范围和精密度等关键性能指标。结果 NM-CLIA检测D1和D2 s Ig E抗体的Lo D均小于0.01 U/m L,线性范围为0.1~100 U/m L。批内精密度小于5%,批间精密度小于8%。方法学比对结果表明,与免疫荧光法相比,屋尘螨过敏原的阳性符合率为95%,阴性符合率为92%,χ~2=174.45,P<0.001,Kappa=0.843,表明NM-CLIA检测屋尘螨过敏原与免疫荧光法有良好的一致性,且±1级符合率为95.6%;粉尘螨过敏原的阳性符合率为91%,阴性符合率为97%,χ~2=154.26,P<0.001,Kappa=0.787,表明NM-CLIA检测粉尘螨过敏原与免疫荧光法有良好的一致性,且±1级符合率为94.2%。结论 NM-CLIA在检测D1和D2 s Ig E抗体时与免疫荧光法有良好的相关性,同时NM-CLIA的Lo D、线性范围和精密度性能优异,可成为D1和D2 s Ig E抗体临床检测方案的推荐方法。

Objective To evaluate the performance of magnetic nanoparticle chemiluminescence immunoassay（NMCLIA） in detection of allergen-specific immunoglobulin E（s Ig E） antibodies to Dermatophagoides pteronyssinus（International Allergen Code D1） and Dermatophagoides farinae（International Allergen Code D2）. Methods A total of 489 serum samples from the patients with suspected allergic disease（244 cases caused by D1, and 245 caused by D2）, who were treated at Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine, were detected by NM-CLIA and immunofluorescence assay, respectively. χ^2 test and Kappa test were used to evaluate the correlation between the two methods in detection of D1 and D2 s Ig E antibodies. The limit of detection（Lo D）, linear range and precision of NM-CLIA in detection of D1 and D2 s Ig E antibodies were verified by the standard method of American Clinical Laboratory Standardization Association. Results The Lo Ds of NM-CLIA in detecting D1 and D2 s Ig E antibodies were both less than 0.01 U/m L, the linearity ranged from 0.1 to 100 U/m L, the within-run precision was less than 5%, and the between-run precision was less than 8%. Methodological comparison results showed that NM-CLIA and immunofluorescence assay had good consistency in detecting D1 and D2 s Ig E antibodies. For D1, the positive coincidence rate and negative coincidence rate were 95% and 92%, respectively（χ^2=174.45, P〈0.001, Kappa=0.843）, and the ±1 class agreement was 95.6%; for D2, the positive coincidence rate and negative coincidence rate were 91% and 97%, respectively（χ^2=154.263,P〈0.001,Kappa=0.787）, and the ±1 class agreement was 94.2%. Conclusion NM-CLIA has good correlation with immunofluorescence assay in detecting D1 and D2 s Ig E antibodies, and has good Lo D, linear range and precision, suggesting that it can be recommended for clinical testing of D1 and D2 s IgE antibodies.