人结核病病原菌型的PCR指印分析

以结核分枝杆菌特异插入序列ＩＳ６１１０两端序列为模板，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ＩＳ６１１０在结核分枝杆菌染色体ＤＮＡ中反复重复且ＩＳ６１１０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

Typing Isolates of Liquid Cultures of TB from Xinjiang by Using the Polymerase Chain Reaction

To develope a molecular typing method for Mycobactereum tuberculases based on the polymerise 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.T his assay utilizes sequenes in IS6110,which occurs as repeated copies in the chromosomes of most M.tuberculoses 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 obtainedinless than 8h,this assay does not requires subculturing,DNA purification,restriction digestion,southern blotting,or nucleic acid hybridization rapid and precise ident}ication of Mtuberculoses strain permits immediate molecular epidemiologic studies.