蛋白芯片技术在自身抗体检测中的应用

Application of ANAs protein chip in autoantibodies profile detection

目的建立应用蛋白芯片技术检测抗核抗体谱中11种自身抗体的方法,并对蛋白芯片技术对自身抗体检测的敏感性和特异性进行评价。方法通过蛋白芯片技术与间接免疫荧光法(IIF)和酶联免疫吸附法(ELISA)自身抗体检测试剂盒进行比对,验证抗核抗体谱检测蛋白芯片(抗SSA-52抗体、抗SSA-60抗体、抗SSB抗体、抗Sm抗体、抗RNP抗体、抗Scl-70抗体、抗Jo-1抗体、抗dsDNA抗体、抗rRNP抗体、抗着丝点抗体和抗核抗体)在临床应用中的敏感性和特异性。除抗Jo-1抗体阳性样品较罕见,仅检测阳性样品32份,其他10种自身抗体,每种选择各阳性样本70份,阴性样本294份;并对检测结果进行统计学分析。结果除抗SSA52抗体和抗SSB抗体的检测敏感度分别为95·7%和98·6%外,其他9种自身抗体的检测敏感度均为100%;除抗SSB抗体、抗RNP-68抗体、抗Scl-70抗体、抗dsDNA抗体、抗CENP-B抗体、抗核抗原提取物抗体(ANA)的检测特异度分别为98·0%、98·0%、99·7%、99·7%、99·7%、98·3%外,其他5种自身抗体的检测特异度均为100%。结论蛋白芯片技术检测11种抗核抗体谱自身抗体的检测敏感度(均>95·0%)和特异度(均>98·0%)均较高,且与IIF和ELISA法有较高的符合率,能满足临床检测自身抗体谱的要求。此外,蛋白芯片技术检测自身抗体时具有高通量、简单快速、敏感度好和特异度强等优点,值得临床推广应用。

Objective To establish and apply the protein chip to detect eleven autoantibodies profile, and evaluate the authenticity and reliability with ANAs protein chip in clinical autoantibodies profile detection. Methods By comparing the results of IIF and ELISA , validation the sensitivity and specificity of ANAs protein chip in clinical autoantibodies profile detection. The autoantibodies detected were anti-SSA- 52, anti-SSA-60, anti-SSB, anti-Sm, anti-RNP, anti-Scl-70, anti-Jo-1, anti-dsDNA, anti-rRNP, anti-centromere antibodies and antinuclear antibodies （ANA）. To each autoantibody, we have selected 70 positive and 294 negative samples except the 32 rare samples that contain anti-Jo-1 antibody. Results The sensitivity to all the autoantibodies was 100% except anti-SSA52 and anti-SSB antibodies was 95.7% and 98. 6% respectively. The specificity to all the autoanbodies was 100% except anti-SSB, anti-RNP-68, anti-Scl-70, anti-dsDNA, anti-CENP-B and ANA was 98. 0%, 98. 0%, 99. 7%, 99. 7%, 99. 7% and 98. 3% respectively Conclusions To all the eleven antinuclear autoantibodies , the sensitivity is all above 95.0% and specificity is all above 98.0%, which indicate that there is high concordances between the ANAs protein chip and the methods used in clinical screening and confirmation, and it could meet the requirement of clinical autoantibodies profile detection. The protein chip method is fast, easy for detection with the characteristic of high-throughput, high sensitivity and specificity, it is hence recommended to apply ANAs protein chip to detect autoantibodies profile in clinical detection.