结核分枝杆菌Rv3871抗原优势肽段的克隆表达及抗原性鉴定

目的:为了提高结核病诊断试剂的特异性和敏感性,克隆表达结核分枝杆菌H37Rv株RD1区Rv3871抗原优势肽段,并应用ELISA法对其抗原性进行初步鉴定。方法:利用Biosun生物信息学软件对Rv3871抗原进行表位分析,通过PCR从结核分枝杆菌H37Rv基因组中扩增Rv3871抗原优势肽段编码基因,在大肠杆菌HB101中进行表达,采用间接ELISA法初步评价其抗原性。结果:Rv3871-1肽段检测的敏感性为32.5%(13/40),特异性为97.2%(35/36);Rv3871-2敏感性为45%(18/40),特异性为100%;Rv3871-3敏感性为37.5%(15/40),特异性为91.6%。结论:结核分枝杆菌RD1区Rv3871抗原优势肽段Rv3871-2有较高的特异性和敏感性,有望作为候选抗原用于结核病患者的血清学检测。

Cloning,Expression and Antigenicity of Rv3871 Dominant Peptide of Mycobacterium tuberculosis

Objective： In order to improve the specificity and sensitivity of diagnostic reagent for the tuberculosis, the Rv3871 dominant peptides in RD1 region of Mycobacterium tuberculosis stain H37Rv were cloned and expressed, and the antigenicity was evaluated by ELISA. Methods： The epitope of Rv3871 antigen was analyzed by using Biosun bioinformatics software. The gene sequences of Rv3871 dominant peptide of H37Rv were cloned by PCR, the genes were induced to express in E.coli HB101 and the antigenicity of the Rv3871 dominant peptides were detected by ELISA. Results： The sensibility of Rv3871-1 peptide was 32.5%（13/40） and the specificity was 97.2%（35/36）; Rv3871-2 peptide had a sensibility of 45%（18/40） with the specificity of 100%; the sensibility of Rv3871-3 peptide was 37.5（15/40） and the specificity was 91.6%（33/36）. Conclusion： The Rv3871-2 dominant peptide of Rv3871 antigen in RD1 region of M.tuberculosis has high specificity and sensitivity, and is expected as a candidate antigen for serological detection of tuberculosis patients.