Abstract：To identify the species of Mycobacterium clinical isolates by molecular biology techniques,six clinical isolates which were preliminarily recognized as Mycobacterium tuberculosis by the TCH and PNB culture methods were selected in this study.PCR was applied to amplify the oxyR-ahpC interval resistant-zone(intergenic region) and the length of PCR product in four strains was the same as that of H37Rv,while the length in the other two strains was different from that of H37Rv but same as Mycobacterium intracellulare 95002.With sequencing and on-line homology comparison with H37Rv,U18263 and U71061,the DNA sequence of oxyR-ahpC intergenic region displayed a 99% homology with Mycobacterium intracellulare U71061 and an 84% homology with Mycobacterium tuberculosis H37Rv.In addition,the results of hsp65 PCR-restriction fragment length polymorphism analysis and multi-locus PCR amplification in the two strains were identical with those of Mycobacterium intracellulare 95002.These two clinical isolates which were preliminarily recognized as Mycobacterium tuberculosis by PNB and TCH culture methods were finally identified as Mycobacterium intracellulare.Results predicted that the application associated with various techniques of molecular biology would provide a faster,easier and more correct way for the species identification of Mycobacterium.
(1)Donnabella V,Salazas SJ,Bonk S,et al.Increasing incidence ofMycobacterium xenopi atBellevue hospital:an emerging patho-gen or a product of improved laboratory methods(J).Chest,2000,118(5);1365-1370.
(2)靳晓红,刘元东,庄玉辉,等.分枝杆菌菌种的基因快速鉴定技术(J).国外医学临床生物化学与检验学分册,2002,23(2):89-90.
(3)Embden JD,Cave MD,Crawford JT,et al.Strain identificationofMycobacterium tuberculosis byDNA fingerprinting:Recom-mendations for a standardized methodology(J).Clin Microbiol,1993,31(8):446-2450.
(4)Huard RC,Lazzarini LC,Butler WR,et al.PCR-Based Methodto Differentiate the Subspecies of theMycobacterium tuberculosiscomplex on the basis of genomic deletions(J).Journal of ClinicalMicrobiology,2003,41(4):1637-1650.
(5)Brunello F,Ligozzi M,Christelli E,et al..Dentification of 54Mycobacterium speciesby PCR-restriction fragment length poly-morphism analysis of the hsp65 gene(J).Clin Microbiol,2001,39:2799-2806.
(6)Chimara E,Ferrazoli L,Ueky SY,et al.Reliable identification ofmycobacterial species by PCR-restriction enzyme analysis(PRA)-hsp65 in a reference laboratory and elaboration of a se-quence-based extended algorithm of PRA-hsp65 patterns(J).BMC Microbiology,2008:(8);48.
(7)刘金伟,匡铁吉,王全河,等.分枝杆菌鉴定方法的比较研究(J).中华结核和呼吸杂志,2000,22(10):631.
(8)Lesk AM.Introduction to Bioinformatics(M).New York:OxfordUniversity Press,2002:1-61.
(9)Koh W,Kwon OJ,Lee KS.Nontuberculous Mycobacterial pulmo-nary diseases in immunocompetent patients(J).Korean Journalof Radiology,2002,3(3):145-157.
(10)Katoch V M.Infections due tonon-tuberculous mycobacteria(J).Indian J Med Res,2004,120:290-304.
