结核分枝杆菌38 kD重组蛋白的纯化及血清学诊断应用价值的研究

Purification of recombinant 38 kD protein and application in serodiagnosis of tuberculosis

目的纯化获得结核分枝杆菌重组38kD蛋白,并评价其在血清学诊断方面的应用价值。方法应用亲和层析方法纯化结核分枝杆菌重组38kD蛋白,应用ELISA方法检测220例血清中的抗结核抗体。结果结核分枝杆菌重组38kD蛋白以包涵体形式表达,以48名正常人血清OD492+2S为正常限值,57例PPD强阳性血清,47例菌阳结核病人血清和68例菌阴结核病人血清阳性检出率分别为13.8%、97.87%和83.82%。结论结核分枝杆菌38kD重组蛋白以包涵体形式高效表达,具有较强的抗原特异性和免疫反应性。

[Objective] To obtain the recombinant 38 kD protein, and evaluate its value in serodiagnosis of tuberculosis. [Methods] The recombinant 38 kD protein was purified by metal chelation chromatography, and its immunogenicity was analyzed in 220 sera by ELISA. [Result] The recombinant 38 kD protein existed in inclusion bodies of TB bacilli. The positive cutoff value was determined as OD492 ＋2S of 48 sera from PPD-negative healthy people detected by ELISA. Of 57 sera from PPD-positive people, 47 sera from smear-positive tuberculosis patients and 68 sera from smear-negative tuberculosis patients, the positive rates of anti-tuberculosis antibodies were 13.8%, 97.87% and 83.82%, respectively. [Conclusion] The recombinant 38 kD protein could be highly expressed in the form of inclusion bodies in TB bacilli. It had good antigenic specificity and immunoreactivity.