结核杆菌重组基因16kD-38kD融合蛋白诊断结核病的应用价值

Evaluation of the recombinant 16kD-38kD fusion protein of Mycobacterium Tuberculosis for serodiagnostic test

目的:构建结核杆菌16kD-38kD重组融合基因,纯化16kD-38kD融合抗原,进一步评价其在结核病血清学诊断中的应用价值。方法:通过构建重组结核杆菌16kD-38kD融合基因工程菌株,经亲和层析纯化融合蛋白,以此蛋白为抗原,建立金标免疫层析法检测结核病人血清中的特异性抗体,同时与单一的16kD重组蛋白和18kD重组蛋白进行比较。结果:16kD、38kD、16kD-38kD三种重组抗原检测肺结核病人血清201份检出率分别56.2%(113/201)、57.7%(116/201)、73.6%(148/201),融合蛋白抗原阳性检出率明显高于单一重组蛋白抗原,对菌阳血清的检出率达到81.9%;检测健康体检血清179份阴性符合率分别为87.2%(156/179)、89.9%(161/179)、88.8%(159/179),三种蛋白无明显差异。结论:16kD-38kD融合蛋白具有较好的敏感性和特异性,与单一使用两种蛋白比较,对结核病人血清抗体的检出率有很大的提高,在结核病血清学诊断中有较高的参考价值。

Objective：To construct the prokaryotic expression vector for gene encoding the 16kD-38kD fusion protein in order to search a serological method to distinguish the state of tuberculous infection.Methods： To gain the recombinant protein antigen 16kD-38kD of Mycobacterium tuberculosis highly expressed in E.coli the fusion protein were purified with affinity chromatography.To build Colloidal gold immunochromatographic assay with the fusion protein antigen in order to detect serum of TB patients.At the same time compare the result with the single 16kD and 38kprotein antigen.Results： 201 patients serum were detect with 16kD,38kD,16kD-38kD three recombinant antigens,the positive rate respectively 56.2%（113/201）,57.7%（116/201）,73.6%（148/201）,the positive rate of 16kD-38kD protein antigen was higher.179 control samples negative rate respectively 87.2%（156/179）,89.9%（161/179）,88.8%（159/179）,shows no evident differences among them.Conclusion： 16kD-38kD fusion protein has good sensitivity and specificity compare with the single 16kD and 38kD protein antigen.In TB serodiagnostic to be higher in reference value.