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重组表达的着丝粒蛋白-B N-端抗原在抗着丝粒自身免疫病血清检测中的应用
APPLICATION OF RECOMBINANT FUSION PROTEIN OF N-TERMINAL FRAGMENT OF CENP-B AS AN ANTIGEN ON THE DETECTION OF SERA WITH ANTI-CENTROMERE AUTOANTIBODY
【摘要】 目的 分析重组表达的CENP B抗原对抗着丝粒自身免疫病血清的鉴定结果 ,探讨其作为临床诊断指标的可行性。 方法 通过构建原核表达载体 ,在大肠杆菌中高效表达CENP BN端 30 0个氨基酸的 1段多肽 ;纯化包涵体 ,制备重组表达的CENP B抗原 ,并利用ELISA、Westernblot的方法对临床诊断为阳性的ACA血清进行鉴定。 结果 重组表达的CENP BN 端抗原对ACA血清的ELISA阳性检出率为 99% ,Westernblot阳性检出准确率为 97%。 结论 重组表达的CENP B抗原可以作为抗着丝粒自身免疫病血清临床诊断的 1个指标。
【Abstract】 Objective Analyzing the detection effects of immune disease sera with ACA (anti-centromere autoantibody) by recombinant fusion protein of CENP-B[a kind of important structural protein in centromere,acting as one of the main antigens of ACA which exist in sera from patients with a variety of progressive systematic sclerosis(PSS) or other auto-immunity disease]. Methods Prokaryon expression vector was constructed,a peptide including 300 amino acids of N-terminal fragment of CENP-B was expressed in Escherichia coli,the recombinant fusion CENP-B antigen was obtained by purifying inclusion bodies.The antigen was used in detection of ACA which shows positive in clinical diagnosis by ELISA and Western blot. Results The positive rate of ACA was 99% by ELISA and 97% by Western blot respectively.Conclusion The recombinant fusion CENP-B antigen may be used as an index in clinical diagnosis for immune disease sera with ACA.
【Key words】 CREST syndrome; CENP-B; Anti-centromere autoantibody (ACA); ELISA; Western blot;
- 【文献出处】 解剖学报 ,Acta Anatomica Sinica , 编辑部邮箱 ,2004年01期
- 【分类号】R593.2
- 【被引频次】4
- 【下载频次】136