结核分枝杆菌16KD重组蛋白的纯化及血清学诊断应用价值的研究

The purification of recombinant 16KD protein and its application in serodiagnosis of tuberculosis

目的：纯化获得结核分枝杆菌重组16KD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16KD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16KD蛋白以包涵体形式表达，以48名正常人血清0D492＋2S为正常限值，57例PPD阳性血清，47例菌阳结核患者血清和68例菌阴结核患者血清阳性检出率分别为19．30％、93．62％和82．35％。结论：结核分枝杆菌16KD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 16KDa protein and evaluate its value in serodiagnosis of tuberculosis. Methods The recombinant 16KD protein was purified by metal chelation chromatography, and its immunogenicity was analyzed in 220 sera by ELISA, Result The recombinant 16KD protein existed in inclusion bodies of E. Coli, 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 19, 30% , 93, 62% and 82. 35%, respectively. Conclusion The recombinant 16KD protein could be highly expressed in the form of inclusion bodies in E. Coli, which had good antigenic specificity and immunoreactivity.