【摘要】 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更多还原

【Abstract】 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 utilizes sequences in IS6110,which occurs as repeated copies in the chromosomes of most M.tuberculosis isolates and produce numerous bands constituting a fingerprint.57 sputums samples strains examined from Xinjiang showed that IS6110 direct PCR is highly discriminatory and reproducible.PCR fingerprinting of the 57 strains generated 7 distinct PCR fingerpriting types,PCR fingerprinting are 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 strain permits immediate molecular epidemiologic studies更多还原