应用生物素-链霉亲合素放大ELISA系统检测抗环瓜氨酸肽抗体  

Determination of anti-CCP antibody by biotin-streptavidin amplified ELISA system

作　　者：房国祥[1,2] 吴云娟[1,2] 王书侠[1,2] 虞伟[2] 李晓军[2] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212001 [2]南京军区南京总医院解放军临床检验医学研究所,南京210002

出　　处：《临床检验杂志》2009年第1期20-23,共4页Chinese Journal of Clinical Laboratory Science

基　　金：中国博士后科学基金资助项目(20060400940);江苏省博士后科学基金资助项目(0602008C);南京军区南京总医院重点课题(Z2008056)

摘　　要：目的用生物素-链霉亲合素放大ELISA系统建立一种敏感、特异、经济简便的抗环瓜氨酸肽(CCP)抗体的测定法。方法用链霉亲合素(SA)预包被,使生物素化的CCP(bio-CCP)固相化,摸索检测抗CCP抗体的最优实验条件,并进行方法学考核和临床应用评价。结果本法对类风湿关节炎(RA)的诊断敏感性为69.4%,特异性为96.8%,阳性预测值为89.1%,阴性预测值为94.1%,阳性似然比为19.28,阴性似然比为0.317,Youden指数为0.658。与欧蒙试剂比对,两法所测抗CCP抗体吸光度/cutoff比值显著相关(n=61,r=0.9253,P<0.001)。在所统计的168例血清样本中,两法同时阳性者60例,同时阴性者105例,一致性百分率达98.2%,结果具有高度一致性(Kappa=0.961,P<0.001)。ROC曲线分析结果显示本法所测抗CCP抗体对RA诊断准确性达到了一个较高水平(AUCROC值=0.901)。在13例诊断为早期RA者中,有10例抗CCP抗体阳性(阳性率76.9%),未发现抗CCP抗体与RA患者临床症状、疾病活动指标(ESR)及RF水平有关。结论成功将生物素-链霉亲合素ELISA系统应用于抗CCP抗体检测,方法敏感、特异、可靠,可用于RA的常规实验室诊断。Objective To develop a highly sensitive, specific and simple enzyme linked immunosorbent assay （ELISA） for determination of anti-cyclic eitrullinated peptide （CCP） antibody. Methods By using the pattern of streptavidin pro-immobilization and conjugation with biotin-CCP （ SA/Bio-CCP）, the experimental conditions of ELISA for anti-CCP antibody were optimized and its methodology and results of clinical application were evaluated. Results The diagnostic sensitivity, specificity, positive/negative predictive value, positive/negative likelihood ratio and Youdeng index in rheumatoid arthritis （RA） patients were 69.4% ,96.8% ,89.1%/94.1% ,19.28/ 0.317 and 0. 658 respectively. Parallel investigation showed that absorbanee and cut-off ratio of anti-CCP antibody was significantly cor- related with Euroimmun Kit（ n = 61,R = 0. 92531,P 〈 0.001 ）. Among 168 samples ,60 positive and 105 negative results were consistent by both assays,and the consistent rate was up to 98.2%. A high consistency with Euroimmun assay（ Kappa =0. 961 ,P 〈0. 001 ） was shown. Area under receiver operative characteristic curve, i. e. AUCROC was 0.901, which suggested that the diagnostic accuracy of anti-CCP antibody tested by the established method was up to a higher level. Of 13 early RA eases, 10 cases were positive （positive rate： 76.9% ）. No association between anti-CCP antibody and clinical symptoms, activity index （ESR） and rheumatoid factor level in RA patients. Conclusion The amplified system of biotin-streptavidin was successfully applied in ELISA for determination of anti-CCP antibody. Its results were sensitive, specific and reliable, so it is suitable for diagnosis of RA in routine laboratories.

关 键 词：环瓜氨酸肽 类风湿关节炎 酶联免疫吸附试验 生物素一链霉亲合素 

分 类 号：R446.6[医药卫生—诊断学]