结核分枝杆菌重组38kD蛋白用于新疆南疆维吾尔族和湖南湘中汉族血清学诊断的研究

The Serodiagnosis of Recombinant 38kD Protein from Mycobacterium Tuberculosis between the Uygur nationality from the South of Sinkiang and the Nan Nationality from the Middle of Hunan

目的:对结核分枝杆菌38kD蛋白编码基因进行克隆表达及纯化,建立基于重组38 kD蛋白的酶联免疫吸附法(ELISA)检测结核病人血清标本,评价重组38 kD蛋白用于结核病血清学诊断抗原的价值。并比较分析其在汉族和维吾尔族人群中的血清学诊断的差异。方法:用PCR方法扩增38 kD蛋白的编码基因,构建重组质粒,转化到大肠杆菌BL21中,经IPTG诱导表达,得到纯化的38 kD蛋白,建立以38 kD蛋白为包被抗原的ELISA,并检测临床确诊的结核病人血清标本。结果:ELISA检测结核病患者血清标本的维吾尔族阳性率为34%(52/153),汉族为52.4%(65/124),两者对比有统计学差异(X2=9.538,P<0.005)。在阴性对照中的维吾尔族特异度为96.4%(159/165),汉族为98.8%(130/133),结果无统计学意义(X2=0.111,P>0.5)。结论:重组38kD蛋白用于血清学诊断的敏感度在维吾尔族和汉族中有差异,而其诊断特异度无差别。

Objective： The encoding gene 38 kD protein of Mycobacterium tuberculosis was cloned, expressed and purified. Established the ELISA assay with recombinant 38kD protein for detecting the specific antibodies in serum samples of patients and evaluating the value of recombinant 38kD protein for diagnose the tuberculosis. Analysis the variability was analyzed by comparing the Han nationality to the Uygur nationality. Methods： Amplify the piece of genes coding 38 kD by PCR was amplified, Recombinant plasmid was constructed, then transformed into E coli BL21 strain, and induced by IPTG. The purified 38KD protein was obtained and established ELISA assay with 38kD protein as the coating antigen and used to detecting blood serum from the TB patients. Results： The positive rate of Uygurnationality is 34%（52/153）, and Han nationality is 52.4%（65/124）, the result have statistical significance.The specificity of Uygur nationality is 96.4%（159/165）, and Han nationality is 98.8%（130/133）, the result has not statistical significance. Conclusion： The sensitivity of recombinant 38 kD protein using serodiagnosis have statistical significance compare the Han nationality to the Uygur nationality and the specificity haven＇t difference.