PCR指印技术对新疆结核病病原Bacter460液体培养标本进行菌株分型

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

THE MOLECULAR TYPING FOR ISOLATES IN LIQUID MEDIA FROM TUBERCULOUS PATIENTS IN XINJIANG BY PCR

To develope a molecular typing method for Mycobacterium tuberculosis based on the polymerase chain reaction, outward-directed PCR primers were designed to the ends of the insertion sequence IS6110.In an attempt to amplify DNA between clusters of this element on the genome, this assay utilized sequences in IS6110, which occurs as repeated copies in the chromosomes of most M.tuberculosis isolates and produce numerous bands Constituting a fingerprint. 31 strains cultured in liquid media from patients in Xinjiang. showed that IS6110-direct PCR was highly discriminatory and reproducible.6 distinct PCR fingerprinting types were found in these 31 strains. PCR fingerprinting was obtained in less than 8h, this assay does not requires subculturing, DNA purification, restriction digestion, Southern blotting, or nucleic-acid hybridization, rapid and precise identification of M.tuberculosis is possible and permits for molecular epidemiologic studies.