液流芯片法在诊断抗中性粒细胞胞浆抗体相关性血管炎中的价值

Role of multiplexed bead-based immunoassay in the determination of AAV

目的探讨液流芯片法在诊断抗中性粒细胞胞浆抗体相关性血管炎(AAV)中的价值。方法选取AAV患者300例,非AAV自身免疫性疾病患者275例及健康对照者277名,分别采用线性免疫印迹法(LIA)和液相芯片法检测抗髓过氧化物酶(MPO)抗体和抗蛋白酶3(PR3)抗体,同时采用间接免疫荧光法(IFA)检测抗中性粒细胞胞浆抗体(ANCA),比较不同检测方法的性能,并筛选诊断AAV的最佳检测方案。结果LIA与液流芯片法检测抗MPO抗体、抗PR3抗体总阳性率较为一致,分别为83.1%、78.2%,kappa值均>0.75。液流芯片法的敏感性和特异性分别为85.7%、82.6%,均高于LIA的59.7%、56.6%(P<0.05)。胞浆型抗中性粒细胞胞浆抗体(cANCA)+抗PR3抗体、核周型抗中性粒细胞胞浆抗体(pANCA)+抗MPO抗体联合检测模式在AAV组中的阳性率最高,分别为42.4%、45.0%。结论液流芯片法对ANCA的检测有较高的敏感性和特异性,与IFA联合检测是诊断AAV的最佳方案。

Objective To investigate the role of multiplexed bead-based immunoassay in the determination of anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis(AAV).Methods A total of 300 patients with AAV,275 autoimmune disease patients without AAV and 277 healthy subjects(healthy control group)were enrolled.The roles of indirect immunofluorescence assay(IFA),line immunoassay(LIA)and multiplexed beadbased immunoassay were evaluated by comparing the results of ANCA,anti-myeloperoxidase(MPO)antibody and anti-protease 3(PR3)antibody determinations.The optimal method for the determination of AAV was screened.Results There was no difference in the positive rate between LIA and multiplexed bead-based immunoassay by comparing the results of anti-MPO antibody and anti-PR3 antibody determinations.The positive rates were 83.1%and 78.2%,respectively,and the kappa value was>0.75.Multiplexed bead-based immunoassay showed higher sensitivity and specificity(85.7%and 82.6%)than those of LIA(59.7%and 56.6%)(P<0.05).The models of cytoplasmic antineutrophil cytoplasmic antibody(cANCA)+PR3 and perinuclear antineutrophil cytoplasmic antibody(pANCA)+MPO showed higher positive rates in the diagnosis of AAV,and the positive rates were 42.4%and 45.0%,respectively.Conclusions Multiplexed bead-based immunoassay shows high sensitivity and specificity for ANCA determination,and the combined determination with IFA plays a role in the diagnosis of AAV.