以结核分枝杆菌38kD重组蛋白为抗原的血清学诊断

Serological Diagnosis Using Recombinant 38 kD Protein of Mycobacterium tuberculosis as Antigen

目的克隆表达结核分枝杆菌38kD抗原基因,并以此蛋白为抗原,进行分枝杆菌的血清学诊断。方法采用PCR自结核分枝杆菌基因组DNA中扩增38kD抗原基因,经测序鉴定正确后,克隆于pGEX-4T-2表达载体,转化大肠杆菌BL21进行诱导表达,利用GSTrapFF蛋白柱纯化表达产物,经ELISA检测其抗原的灵敏度与特异性。结果目的蛋白表达量约占菌体总蛋白量的28%,ELISA检测灵敏度为79.8%,特异性为99.0%。结论重组38kD抗原可用于结核分枝杆菌的诊断。

Objective To clone and express the gene encoding 38 kD protein of Mycobacterium tuberculosis and perform serological diagnosis using the protein as an antigen. Methods Amplify 38 kD antigen gene from the genomic DNA of Mycobacterium tuberculosis by PCR,identify by sequencing,then clone into expression vector pGEX-4T-2 and transform to E. coli BL21 for expression. Purify the expressed product by GSTrapFF protein column chromatography and test for sensitivity and specificity by ELISA. Results The expressed product contained 28% of total somatic protein ,and its sensitivity and specificity were 79. 8% and 99. 0% respectively. Conclusion The recombinant 38 kD protein might be used as an antigen for diagnosis of infection with Mycobacterium tuberculosis.