蛋白芯片法检测抗可提取性核抗原抗体  

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

作　　者：温晓宏[1] 王玉华[1] 杨玲[2] 吴莹[1] 张奉春[1] 曾小峰[1] 赵岩[1] 

机构地区：[1]中国医学科学院中国协和医科大学北京协和医院风湿免疫科,100730 [2]北京华大基因研究中心诊断试剂部

出　　处：《中国药物与临床》2007年第6期405-407,共3页Chinese Remedies & Clinics

基　　金：国家自然科学基金资助项目(30300164)

摘　　要：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。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-Sin （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 （k=0.78）, anti-Scl-70 （K=0.83） and anti-Jo-1 （k=1.00） （all k〉0.75, P〈0.05）; ③ head-to-head comparison using the 1580 consecutive samples showed that both methods were well consistent （k〉0.75） in detection of anti-RNP （k=0.83）,anti-SCL-70 （k=0.79） and anti-Jo-1 （k=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.

关 键 词：蛋白质阵列分析 免疫扩散 抗可提取性核抗原抗体 

分 类 号：R446.6[医药卫生—诊断学]