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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。
OBJECTIVE: To improve the specificity and sensitivity of the diagnostic reagent for tuberculosis, the dominant epitope of Rv3871 antigen in RD1 region of Mycobacterium tuberculosis strain H37Rv was cloned and its antigenicity was preliminarily identified by ELISA. Methods: The epitope of Rv3871 antigen was analyzed by Biosun bioinformatics software. The gene encoding the dominant peptide of Rv3871 antigen was amplified by PCR from Mycobacterium tuberculosis H37Rv genome and expressed in E. coli HB101. The indirect ELISA was used to evaluate the epitope of Rv3871 antigen. Its antigenicity. Results: The sensitivity of Rv3871-1 was 32.5% (13/40) and the specificity was 97.2% (35/36). The sensitivity of Rv3871-2 was 45% (18/40) and the specificity was 100% The sensitivity of Rv3871-3 was 37.5% (15/40) and the specificity was 91.6%. Conclusion: The Rv3871 antigen of Rb3871-2 in Mycobacterium tuberculosis RD1 is highly specific and sensitive and may be used as a candidate antigen in the serological detection of tuberculosis patients.