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    • 33. 发明申请
    • A METHOD FOR THE DIAGNOSIS, IDENTIFICATION AND CHARACTERIZATION OF M. TUBERCULOSIS AND OTHER MYCOBACTERIA BY SHIFT MOBILITY ASSAY
    • 一种通过移动移动测定法诊断,鉴定和表征M.TUBERCULOSIS和其他MYCOBACTERIA的方法
    • WO1999035284A1
    • 1999-07-15
    • PCT/BR1997000087
    • 1997-12-30
    • UNIVERSIDADE FEDERAL DE MINAS GERAISPEREGRINO FERREIRA, Paulo, CésarGEESSIEN KROON, ErnaBERNARDES MARGUTTI PINTO, Maria, ElizabethALEIXO, Agdemir, Wáleria
    • UNIVERSIDADE FEDERAL DE MINAS GERAIS
    • C12Q01/68
    • C12Q1/68C12Q1/6806C12Q1/689
    • The present invention relates to a method for diagnosis, identification and characterization of M. tuberculosis or any other mycobacteria by using PCR and shift mobility assay (SMA). The method is based both on microheterogeneities observed within the 16S rRNA sequences established for mycobacteria and on nucleotide gaps and mismatches which cause a decrease in the electrophoretic mobility of DNA heteroduplex in polyacrylamide gels. The PCR strategy is based on the use of a specific primers for mycobacteria genera and divergence in sequences found in 16S rRNA coding gene. The difference in nucleotide sequence was enough to identify mycobacteria species, since a remarkable shift between single stranded and homoduplex bands in PAGE were observed among mycobacteria tested, when Mycobacterium tuberculosis was used as standard. The shift of PCR product of bacteria other than mycobacteria was observed above the single stranded band in PAGE, both in standard cultures and clinical specimens. SMA can thus provide a fast and sensitive method for detection and classification of mycobacteria in clinical samples as well as pure culture.
    • 本发明涉及通过使用PCR和移位移动性测定(SMA)来诊断,鉴定和表征结核分枝杆菌或任何其它分枝杆菌的方法。 该方法基于在为分枝杆菌建立的16S rRNA序列中观察到的微观异质性以及导致聚丙烯酰胺凝胶中DNA异源双链的电泳迁移率降低的核苷酸间隙和错配。 PCR策略是基于使用特异性引物分枝杆菌属和16S rRNA编码基因中发现的序列发散。 核苷酸序列的差异足以识别分枝杆菌物种,因为当使用结核分枝杆菌标准时,在测试的分枝杆菌中观察到在单链和同源双链条之间的显着变化。 在标准培养物和临床标本中,在单链条带上均观察到除分枝杆菌以外的细菌的PCR产物位移。 因此,SMA可以提供用于临床样品中的分枝杆菌的检测和分类以及纯培养的快速和灵敏的方法。