国际免疫学杂志
國際免疫學雜誌
국제면역학잡지
INTERNATIONAL JOURNAL OF IMMUNOLOGY
2014年
6期
520-525
,共6页
类风湿关节炎%滑膜组织%cDNA文库%SEREX技术%自身抗原
類風濕關節炎%滑膜組織%cDNA文庫%SEREX技術%自身抗原
류풍습관절염%활막조직%cDNA문고%SEREX기술%자신항원
Rheumatoid arthritis%Synovium%cDNA library%SEREX technology%Self-antigen
目的 通过SEREX(serological analysis of recombinant cDNA expression library)技术对类风湿关节炎滑膜组织cDNA文库进行初步筛选,得到新的类风湿关节炎自身抗原.方法 利用SMART技术构建类风湿关节炎患者滑膜组织cDNA文库,并对该文库进行SEREX筛选,获得特异性结合阳性克隆,对阳性克隆同源性分析.然后进行酶联免疫吸附实验(ELISA)验证,分析该阳性克隆对于类风湿关节炎诊断临床敏感度和特异度.结果 构建cDNA文库滴度约为6.89×106 pfu/mL,筛选文库获得12个阳性克隆,同源性分析3个阳性克隆插入片段为已知蛋白,分别是:Filaggrin、SERPINE2和p68.进行血清学初步验证,抗-Filaggrin和抗-SERPINE2具有高敏感度和低特异度,而抗-p68具有高敏感度和高特异度.结论 抗-Filaggrin、抗-SERPINE2和抗-p68对于类风湿关节炎的诊断具有不同的敏感度和特异度.
目的 通過SEREX(serological analysis of recombinant cDNA expression library)技術對類風濕關節炎滑膜組織cDNA文庫進行初步篩選,得到新的類風濕關節炎自身抗原.方法 利用SMART技術構建類風濕關節炎患者滑膜組織cDNA文庫,併對該文庫進行SEREX篩選,穫得特異性結閤暘性剋隆,對暘性剋隆同源性分析.然後進行酶聯免疫吸附實驗(ELISA)驗證,分析該暘性剋隆對于類風濕關節炎診斷臨床敏感度和特異度.結果 構建cDNA文庫滴度約為6.89×106 pfu/mL,篩選文庫穫得12箇暘性剋隆,同源性分析3箇暘性剋隆插入片段為已知蛋白,分彆是:Filaggrin、SERPINE2和p68.進行血清學初步驗證,抗-Filaggrin和抗-SERPINE2具有高敏感度和低特異度,而抗-p68具有高敏感度和高特異度.結論 抗-Filaggrin、抗-SERPINE2和抗-p68對于類風濕關節炎的診斷具有不同的敏感度和特異度.
목적 통과SEREX(serological analysis of recombinant cDNA expression library)기술대류풍습관절염활막조직cDNA문고진행초보사선,득도신적류풍습관절염자신항원.방법 이용SMART기술구건류풍습관절염환자활막조직cDNA문고,병대해문고진행SEREX사선,획득특이성결합양성극륭,대양성극륭동원성분석.연후진행매련면역흡부실험(ELISA)험증,분석해양성극륭대우류풍습관절염진단림상민감도화특이도.결과 구건cDNA문고적도약위6.89×106 pfu/mL,사선문고획득12개양성극륭,동원성분석3개양성극륭삽입편단위이지단백,분별시:Filaggrin、SERPINE2화p68.진행혈청학초보험증,항-Filaggrin화항-SERPINE2구유고민감도화저특이도,이항-p68구유고민감도화고특이도.결론 항-Filaggrin、항-SERPINE2화항-p68대우류풍습관절염적진단구유불동적민감도화특이도.
Objective To obtain new RA antigen genes,we screened the cDNA library of RA Using SEREX technology.Methods We constructed the cDNA library using SMART technology.After evaluation,the cDNA library was screened with the RA sera.The positive clones were obtained and the nucleotide sequences of positive clones were identified and analyzed with BLAST software in GenBank.According to the results of ELISA,we analysed the specificity and sensitivity for the diagnosis and treatment of RA.Results The titer of cDNA library was 6.89 × 106 pfu/mL.After several rounds SEREX screening,12 positive clones were obtained.3 of 12 genes were homologous to the genes known in GenBank.They are Filaggrin,SERPINE2 and p68.The results of ELISA were as follows.Anti-Filaggrin and anti-SERPINE2 are of high sensitivity but low specificity; While,anti-p68 is of high sensitivity and specifity.Conclusion Anti-Filaggrin、anti-SERPINE2 and anti-p68 are of different sensitivity and specifity for diagnosis of RA.
