结核分枝杆菌CFP10ESAT6重组融合蛋白抗原血清学诊断价值

Serodiagnostic value of recombinant CFP10ESAT6 protein from Mycobacterium tuberculosis

摘要:

目的研究重组CFP10ESAT6蛋白的抗原性，评价其在结核病血清学诊断中的价值，探寻更有效的结核病诊断试剂。方法应用基因工程技术表达、纯化rCFP10ESAT6蛋白，通过酶联免疫吸附试验检测192例健康者和210例结核病患者血清与纯化rCFP10ESAT6的抗体反应。结果 rCFP10ESAT6蛋白在大肠埃希菌细胞内以包涵体形式高效表达，表达量约占菌体总蛋白的25％，分子量约为28 kD，具有良好的抗原特异性。在210例结核病患者血清中，103例菌阳患者和107例菌阴患者抗rCFP10ESAT6抗体的阳性率分别为30.10％(31/103)和28.97％(31/107)，总的灵敏度为29.52%(62/210)；在192例健康者血清中，95例PPD阴性者和97例PPD阳性者抗rCFP10ESAT6抗体的阳性率分别为2.11%(2/95)和6.19%(6/97)，总的特异性为95.83%(184/192)。结论rCFP10ESAT6蛋白可能成为结核病血清学诊断的组合抗原之一。

关键词:

结核;结核分枝杆菌;rCFP10ESAT6;诊断;实验室技术和方法

Abstract:

ObjectiveTo evaluate the value of recombinant culture filter protein10early secretory antigenic target 6 （rCFP10ESAT6） protein from Mycobacterium tuberculosis (M. tuberculosis) on the serodiagnosis of tuberculosis (TB), and to develop more effective diagnostic reagents for tuberculosis. MethodsThe rCFP10ESAT6 protein was expressed and purified by metal chelation chromatography. Sera of 192 healthy persons and 210 TB patients were detected the antibodies against M. tuberculosis rCFP10ESAT6 antigens by ELISA. ResultsThe rCFP10ESAT6 protein existed in inclusion bodies of Escherichia coli, and amounted to 25% of total bacterial protein, its molecular weight was estimated about 28 kD. Of sera from 210 TB patients, the sensitivity of antibody detection using rCFP10ESAT6 protein was 30.10% (31/103) and 28.97％ (31/107) in 103 bacteriumpositive patients and 107 bacteriumnegative patients, overall sensitivity was 29.52% (62/210);of 192 healthy subjects, the sensitivity of antibody detection using rCFP10ESAT6 protein was 2.11% (2/95) in 95 PPDnegative persons and 6.19% (6/97) in 97 PPDpositive persons, respectively, overall specificity was 95.83% (184/192).ConclusionThe recombinant CFP10ESAT6 might be used as one of multiple antigens in the serodiagnosis of tuberculosis.

Key words:

tuberculosis; Mycobacterium tuberculosis; rCFP10ESAT6; diagnosis; laboratory technique and methods