Rapid identification of mycobacteria by gene amplification restriction analysis technique targeting 16S-23S ribosomal RNA internal transcribed spacer & flanking region.

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dc.contributor.authorKatoch, V Men_US
dc.contributor.authorParashar, Deeptien_US
dc.contributor.authorChauhan, D Sen_US
dc.contributor.authorSingh, Den_US
dc.contributor.authorSharma, V Den_US
dc.contributor.authorGhosh, Sen_US
dc.date.accessioned2007-02-14en_US
dc.date.accessioned2009-05-27T07:00:04Z
dc.date.available2007-02-14en_US
dc.date.available2009-05-27T07:00:04Z
dc.date.issued2007-02-14en_US
dc.description.abstractBACKGROUND & OBJECTIVE: Conventional identification of a clinical isolate of mycobacteria primarily based on culture characteristics and biochemical tests needs several weeks and may remain inconclusive. This study was undertaken to develop a new rapid method to identify the mycobacterial isolates at species level by gene amplification restriction analysis using primers encoding 16S-23S rRNA internal transcribed spacer (ITS) region and flanking parts of the 16S as well as 23S rRNA gene. METHODS: This system is based on the amplification of approximately 1.8 kb fragment encoding 16S-23S rRNA spacer region and flanking parts of the 16S as well as 23S rRNA gene. This assay was applied on 13 reference strains and 480 clinical isolates of mycobacteria to validate the technique. Restriction was carried out with three restriction endonucleases Hha I, Hinf I and Rsa I. RESULTS: Distinct gene amplification restriction analysis patterns were obtained by restriction of amplicons with three distinct restriction endonucleases (Hha I, Hinf I and Rsa I) which could differentiate various mycobacterial species. INTERPRETATION & CONCLUSION: Restriction patterns with the enzymes used in this study could clearly distinguish Mycobacterium tuberculosis complex from other non chromogenic clinically important species M. avium and M. intracellulare. Results indicated this assay to be a simple, rapid and reproducible method to identify clinically relevant mycobacteria.en_US
dc.description.affiliationDepartment of Microbiology & Molecular Biology, National JALMA Institute for Leprosy & Other Mycobacterial Disease, (ICMR), Taj Ganj, Agra 282001, India. vishwamohan_katoch@yahoo.co.inen_US
dc.identifier.citationKatoch VM, Parashar D, Chauhan DS, Singh D, Sharma VD, Ghosh S. Rapid identification of mycobacteria by gene amplification restriction analysis technique targeting 16S-23S ribosomal RNA internal transcribed spacer & flanking region. Indian Journal of Medical Research. 2007 Feb; 125(2): 155-62en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/18868
dc.language.isoengen_US
dc.source.urihttps://icmr.nic.in/ijmr/ijmr.htmen_US
dc.subject.meshBacterial Typing Techniques --methodsen_US
dc.subject.meshMycobacterium --geneticsen_US
dc.subject.meshNucleic Acid Amplification Techniquesen_US
dc.subject.meshRNA, Ribosomal, 16S --geneticsen_US
dc.subject.meshRNA, Ribosomal, 23S --geneticsen_US
dc.subject.meshRestriction Mappingen_US
dc.titleRapid identification of mycobacteria by gene amplification restriction analysis technique targeting 16S-23S ribosomal RNA internal transcribed spacer & flanking region.en_US
dc.typeComparative Studyen_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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