Zhang L, Zhuang Y, He X, Zhang X, Li G
Tuberculosis Research Laboratory, 309 Hospital of PLA, Beijing 100091.
Wei Sheng Wu Xue Bao. 2000 Oct;40(5):459-64.
The 16S-23S ribosomal DNA spacer sequence of mycobacterium were amplified by PCR. The products were visualized by PAGE, and evaluate the possibility for classification and identification of mycobacterium at gene level. The sensitivity of PCR in annealing temperature 45 degrees C was 500 fg/microL, whereas 50 degrees C was 5 pg/microL. The results showed that: the amplified bands ranging from 300 to 600 bp, most of rapid-growing Mycobacterium and Nonmycobacterium tested have more bands and the bands molecular weights were larger than slow-growing Mycobacterium. The relativity of mycobacterium < 70%, most of them < 50%. This experimental method might be rapid and effective for differentiation of Mycobacterium at species level.
采用聚合酶链反应（PCR）扩增分枝杆菌的16S - 23S核糖体DNA间隔序列。产物通过聚丙烯酰胺凝胶电泳（PAGE）进行可视化，并评估在基因水平上对分枝杆菌进行分类和鉴定的可能性。PCR在退火温度45℃时的灵敏度为500 fg/μL，而在50℃时为5 pg/μL。结果表明：扩增条带范围为300至600 bp，大多数测试的快速生长分枝杆菌和非分枝杆菌有更多条带且条带分子量大于缓慢生长分枝杆菌。分枝杆菌之间的相关性<70%，大多数<50%。该实验方法可能对分枝杆菌的种水平鉴别快速有效。
