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以结核分枝杆菌特异插入序列 IS61 1 0两端序列为模板 ,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 IS61 1 0在结核分枝杆菌染色体DNA中反复重复且 IS61 1 0序列之间相距较近 ,经 PCR扩增呈现多条带型构成 DNA指印。采用 PCR指印技术 ,对人结核病患者痰标本中的结核分枝杆菌进行分型 ,57份痰标本呈现 7种 PCR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。经过对 57份标本的检测 ,证明该法是一种快速、准确的鉴定与分析方法 ,并可直接进行分子流行病学研究
A pair of exo-PCR primers were designed for PCR amplification using the two sequences of Mycobacterium tuberculosis-specific insertion sequence IS6110 as a template to establish a rapid molecular biological typing technique for Mycobacterium tuberculosis. The experiment was based on the use of IS61 10 was repeatedly repeated in M. tuberculosis chromosomal DNA and the IS6110 sequences were close to each other. A number of banding DNA fingerprints were found by PCR amplification. PCR fingerprinting was used to genotype Mycobacterium tuberculosis in sputum samples from patients with tuberculosis. Seven fingerprints were obtained from 57 sputum samples. This PCR typing technique needs a short time for the typing of Mycobacterium tuberculosis and does not require complicated steps such as bacterial culture, DNA purification and digestion, Suther transfer or nucleic acid hybridization. After the detection of 57 specimens, this method is a rapid and accurate identification and analysis method, and can be directly conducted molecular epidemiological studies