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以结核分枝杆菌特异插入序列IS6 110两端序列为模板 ,设计一对外向PCR引物进行PCR扩增 ,从而建立一种结核分枝杆菌的快速分子生物学分型技术 ,该试验的基础是利用IS6 110在结核分枝杆菌染色体DNA中反复重复且IS6 110序列之间相距较近 ,经PCR扩增呈现多条带型构成DNA指印。在对新疆结核病人的 31份液体培养标本的PCR检测呈现 6种指印 ,该PCR分型技术对结核分枝杆菌的分型所需时间短 ,不需细菌再培养 ,DNA纯化和酶切、萨瑟恩转印或核酸杂交等繁琐步骤。证明该法是一种快速、准确的鉴定与分型方法并可直接进行分子流行病学研究
A pair of exogenous PCR primers was designed for PCR amplification using the both ends of IS6 110 sequence of Mycobacterium tuberculosis as a template to establish a rapid molecular biological typing method for Mycobacterium tuberculosis based on the use of IS6 110 was repeatedly repeated in the Mycobacterium tuberculosis chromosomal DNA, and the IS6 110 sequences were close to each other. A number of banding DNA fingerprints were appeared by PCR amplification. Sixty-six fingerprints were detected by PCR in 31 liquid culture samples of tuberculosis patients in Xinjiang. The PCR typing technique required less time for the typing of M. tuberculosis, no bacterial re-culturing, DNA purification and enzyme digestion, Thur transfer or nucleic acid hybridization and other tedious steps. Prove that this method is a rapid and accurate identification and typing method and can be directly conducted molecular epidemiological studies