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采用PCR(聚合酶链反应)技术,选择一对寡核苷酸引物,扩增的靶基因为克隆的PH_重组标准人型结核分枝杆菌2.4kb DNA。其基因片段是结核杆菌所特有,单一拷贝基因,与其他非典型分枝杆菌的基因无同源序列。研究结果表明:①扩增了人型和牛型结核杆菌标准株基因DNA 158bp 片段,而其他14种非典型分枝杆菌和金黄色葡萄球菌、脓杆菌 DNA 未扩增出相应产物.②10倍系列稀释人型结核菌 DNA,检测敏感性相当于10～50菌数/ml痰样。③60例临床肺结核病人痰样进行 PCR 技术检测和常规生物法测定比较,前者阳性率为58.3%,后者检测率11.70%。而20例非结核痰样用两种方法检测全为阴性。提示:PCR 技术诊断结核病快速、简易、特异、敏感、高效,是在基因水平上检测的一种新技术。
A pair of oligonucleotide primers was selected by PCR (Polymerase Chain Reaction) technique and the amplified target gene was cloned PH_Recombinant standard human Mycobacterium tuberculosis 2.4 kb DNA. The gene fragment is unique to Mycobacterium tuberculosis, a single copy of the gene, and no other genes of atypical mycobacterial homology. The results showed that: (1) 158bp DNA was amplified from standard strains of human and mycobacterium tuberculosis, while the other 14 kinds of atypical mycobacteria, Staphylococcus aureus and Pseudomonas aeruginosa did not amplify the corresponding products.②10-fold serial dilutions Human TB DNA, detection sensitivity is equivalent to 10 ~ 50 bacteria / ml sputum samples. ③60 cases of clinical pulmonary tuberculosis sputum samples detected by PCR and conventional biological method, the former positive rate was 58.3%, the latter detection rate of 11.70%. The 20 cases of non-tuberculosis sputum samples detected by both methods were all negative. Tip: PCR technology to diagnose tuberculosis is rapid, simple, specific, sensitive and efficient. It is a new technology detected at the gene level.