蛋白芯片法检测抗可提取性核抗原抗体

目的利用蛋白芯片方法检测抗可提取性核抗原(ENA)抗体,并与免疫双扩散(ID)相比较,评价蛋白芯片方法的敏感度、特异度。方法分3组血清:①抗ENA抗体标准血清10份;②献血员血清200份及诊断明确患者血清445份;③实验室连续常规送检血清1580份。蛋白芯片法:采用华大基因研究中心研究的板式蛋白芯片,测定方法参照其说明书进行;ID:采用经典的梅花孔,所用抗原为人脾提取物。结果①蛋白芯片法测得10份抗ENA抗体标准血清结果全部准确。②献血员200份血清两种方法检测抗ENA抗体全部阴性;患者血清结果显示蛋白芯片法检测抗Sm抗体、抗RNP抗体、抗SSA抗体和抗SSB抗体的敏感度(10.93%、19.73%、56.27%、30.13%)高于ID(5.33%、15.47%、42.40%、10.13%)(P<0.05),抗RNP抗体(κ=0.78)、抗Scl-70抗体(κ=0.83)及抗Jo-1抗体(κ=1.00)在两种方法均有很好一致性,κ>0.75(P<0.05)。③与ID同步盲法比较的1580份血清,抗RNP抗体(κ=0.83)、抗SCL-70抗体(κ=0.79)、抗Jo-1抗体(κ=0.92)三种抗体两种方法有很好的一致性κ>0.75;在抗Sm抗体、抗RNP抗体、抗SSA抗体、抗SSB抗体4种抗体蛋白芯片法敏感度(5.89%、8.40%、29.67%、14.21%)高于ID法(1.65%、6.83%、18.13%、4.95%)(P<0.05)。结论①蛋白芯片法较为敏感、特异,与ID一致性较好。②蛋白芯片法检测抗Sm抗体、抗RNP抗体、抗SSA抗体、抗SSB抗体敏感度高于ID法。

Protein Array Analysis for detection of anti-extractable nuclear antigen antibodies

Objective To evaluate the sensitivity and specificity of protein array analysis vs double immunodiffusion(ID) for detection of anti-extractable nuclear antigen(anti-ENA) antibodies in sera of patients with connective tissue diseases(CTD) .Methods Proteinchips kit with highly purified native antigen(Array-ELISA,Beijing Genomics Institute) and double immunodiffusion(ID) (using the classical 5-well pattern and home-made human spleen extracts as antigen) were employed to detect the anti-ENA antibodies in three groups of serum samples:① 10 of anti-ENA standards;② 200 from healthy blood donors and 451 from confirmed patients with CTD;③ 1580 consecutive samples sent to our laboratory for screening of CTD.Results ① Array-ELISA proved accurate in test of all serum standards;② The serum samples from 200 healthy blood donors tested negative by the both methods.Among 445 patient serum samples,Array-ELISA method showed higher sensitivity for detection of anti-Sm(10.93% vs 5.33%) ,anti-RNP(19.73% vs 15.47%) ,anti-SSA(56.27% vs 42.40%) and anti-SSB(30.13% vs 10.13%) compared with ID method(all P<0.05) ,while the both were well consistent with regard to detection of anti-RNP(κ=0.78) ,anti-Scl-70(κ=0.83) and anti-Jo-1(κ=1.00) (all κ>0.75,P<0.05) ;③ head-to-head comparison using the 1580 consecutive samples showed that both methods were well consistent(κ>0.75) in detection of anti-RNP(κ=0.83) ,anti-SCL-70(κ=0.79) and anti-Jo-1(κ=0.92) ,while Array-ELISA was more sensitive for detection of anti-Sm(5.89% vs 1.65%) ,anti-RPN(8.40% vs 6.83%) ,anti-SSA(29.67% vs 18.13%) and anti-SSB(14.21% vs 4.95%) compared with ID method(all P<0.05) .Conclusion ① Array-ELISA is both sensitive and specific and shows good consistency with ID method.② In testing for anti-Sm,anti-RNP,anti-SSA and anti-SSB,Array-ELISA appears more sensitive compared with ID method.