Zhuang Y, Wu X, Zhang X, Li G
809 Hospital of PLA Tuberculosis Research Laboratory, Beijing.
Wei Sheng Wu Xue Bao. 1992 Oct;32(5):364-9.
The PCR was used to detect M. tuberculosis DNA sequences in uncultured clinical specimens. Two oligonucleotide primers with 20 bp each amplified target template DNA of M. tuberculosis. Amplified DNA product was 245 bp which was identified by agarose gel electrophoresis. The sensitivity of detection of M. tuberculosis genomic DNA and bacteria suspension by PCR was lpg and 13 viable bacteria cell/ml, respectively. In specificity experiments, only M. tuberculosis, M. bovis and BCG were positive by PCR, but all other 14 Mycobacterium tested, including streptomyces lividans and E. coli plasmid PUC19 were negative. The sensitivity of detection of M. tuberculosis by PCR was determined by comparing the fast-acid staining and culture on total 54 sputum specimens of pulmonary tuberculosis and 12 nontuberculosis lung disease. The positive rate of PCR in pulmonary tuberculosis were 37.0%, culture method showed only 14.8%, fast-acid staining were 16.7%. Nontuberculosis lung disease were negative. The results show that DNA amplification is a superior method with high degree of sensitivity and specificity for rapid diagnosis of pulmonary tuberculosis.
采用聚合酶链反应（PCR）检测未经培养的临床标本中的结核分枝杆菌DNA序列。两条各含20个碱基对的寡核苷酸引物扩增结核分枝杆菌的靶模板DNA。扩增的DNA产物为245个碱基对，通过琼脂糖凝胶电泳进行鉴定。PCR检测结核分枝杆菌基因组DNA和细菌悬液的灵敏度分别为1皮克和每毫升13个活细菌细胞。在特异性实验中，仅结核分枝杆菌、牛分枝杆菌和卡介苗经PCR检测呈阳性，而所检测的其他14种分枝杆菌，包括淡紫链霉菌和大肠杆菌质粒PUC19均为阴性。通过对54份肺结核痰标本和12份非结核性肺病痰标本进行抗酸染色和培养，并与PCR检测结果进行比较，确定了PCR检测结核分枝杆菌的灵敏度。肺结核患者中PCR的阳性率为37.0%，培养法仅为14.8%，抗酸染色为16.7%。非结核性肺病患者均为阴性。结果表明，DNA扩增是一种对肺结核快速诊断具有高度敏感性和特异性的优越方法。
