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目的对结核分枝杆菌Rv3873进行生物信息学分析及抗原表位预测,原核表达Rv3873基因21～235位氨基酸蛋白片段,命名为Rv387321～235重组蛋白,初步分析其抗原性。方法 PCR扩增结核分枝杆菌Rv3873 21～235位氨基酸的基因片段,构建其原核表达载体,转化大肠杆菌BL21(DE3),获得重组蛋白并纯化;采用酶联免疫吸附试验(ELISA)对Rv387321～235重组蛋白进行抗原性检测。结果具有优势抗原表位的Rv3873蛋白片段在大肠杆菌中高效表达;ELISA结果显示,Rv387321～235重组蛋白在结核病检测中的敏感性为28%,特异性94.6%。结论生物信息学预测Rv3873基因抗原表位,为寻找有价值的抗原靶点提供了参考;具有优势抗原表位的重组蛋白片段在结核病诊断中具有潜在应用价值,可作为联合诊断的备选抗原之一。
Objective To analyze the bioinformatics analysis and epitope prediction of Mycobacterium tuberculosis Rv3873 and express the Rv387332 ~ 235 recombinant protein by prokaryotic expression, and analyze its antigenicity. Methods The gene fragment of amino acids 21-235 of Mycobacterium tuberculosis Rv3873 was amplified by PCR. The prokaryotic expression vector was constructed and transformed into E. coli BL21 (DE3) to obtain the recombinant protein. The recombinant protein was purified and purified by enzyme linked immunosorbent assay (ELISA) 235 recombinant protein antigenicity test. Results The Rv3873 protein fragment with dominant epitopes was highly expressed in E. coli. The results of ELISA showed that the sensitivity and specificity of Rv387321 ~ 235 recombinant protein in detecting tuberculosis were 28% and 94.6%, respectively. Conclusion Bioinformatics prediction of Rv3873 gene epitope provides a reference for searching for valuable antigenic targets. Recombinant protein fragments with dominant epitopes have potential value in the diagnosis of tuberculosis and can be used as candidate antigen for combined diagnosis one.