Mycobacterial Interspersed Repetitive-Unit-Variable-Number Tandem-Repeat (MIRU-VNTR) Genotyping of Mycobacterium intracellulare for Strain Comparison with Establishment of a PCR-Based Database

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dc.contributorVirginia Techen
dc.contributor.authorIakhiaeva, Elenaen
dc.contributor.authorMcNulty, Stevenen
dc.contributor.authorBrown-Elliott, Barbara A.en
dc.contributor.authorFalkinham, Joseph O. IIIen
dc.contributor.authorWilliams, Myra D.en
dc.contributor.authorVasireddy, Ravikiranen
dc.contributor.authorWilson, Rebecca W.en
dc.contributor.authorTurenne, Christineen
dc.contributor.authorWallace, Richard J. Jr.en
dc.contributor.departmentBiological Sciencesen
dc.date.accessed2015-11-28en
dc.date.accessioned2015-11-29T04:13:27Zen
dc.date.available2015-11-29T04:13:27Zen
dc.date.issued2012-11-21en
dc.description.abstractStrain comparison is important to population genetics and to evaluate relapses in patients with Mycobacterium avium complex (MAC) lung disease, but the "gold standard" of pulsed-field gel electrophoresis (PFGE) is time-consuming and complex. We used variable-number tandem repeats (VNTR) for fingerprinting of respiratory isolates of M. intracellulare from patients with underlying bronchiectasis, to establish a nonsequence-based database for population analysis. Different genotypes identified by PFGE underwent species identification using a 16S rRNA gene multiplex PCR. Genotypes of M. intracellulare were confirmed by internal transcribed spacer 1 (ITS1) sequencing and characterized using seven VNTR primers. The pattern of VNTR amplicon sizes and repeat number defined each specific VNTR type. Forty-two VNTR types were identified among 84 genotypes. PFGE revealed most isolates with the same VNTR type to be clonal or exhibit similar grouping of bands. Repetitive sequence-based PCR (rep-PCR) showed minimal pattern diversity between VNTR types compared to PFGE. Fingerprinting of relapse isolates from 31 treated patients using VNTR combined with 16S multiplex PCR unambiguously and reliably distinguished different genotypes from the same patient, with results comparable to those of PFGE. VNTR for strain comparison is easier and faster than PFGE, is as accurate as PFGE, and does not require sequencing. Starting with a collection of 167 M. intracellulare isolates, VNTR distinguished M. intracellulare into 42 clonal groups. Comparison of isolates from different geographic areas, habitats, and clinical settings is now possible.en
dc.description.sponsorshipAmon G. Carter Foundationen
dc.description.sponsorshipNTM-IR private foundationen
dc.description.sponsorshipThe University of Texas Health Science Center at Tyleren
dc.description.sponsorshipVirginia Tech. Department of Biological Sciencesen
dc.description.sponsorshipSaskatchewan Disease Control Laboratoryen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationIakhiaeva, Elena et al. (2013). Mycobacterial Interspersed Repetitive-Unit-Variable-Number Tandem-Repeat (MIRU-VNTR) Genotyping of Mycobacterium intracellulare for Strain Comparison with Establishment of a PCR-Based Database. Journal of Clinical Microbiology, 51(2), 409-416. doi:10.1128/jcm.02443-12en
dc.identifier.doihttps://doi.org/10.1128/jcm.02443-12en
dc.identifier.issn0095-1137en
dc.identifier.urihttp://hdl.handle.net/10919/64224en
dc.identifier.urlhttp://jcm.asm.org/content/51/2/409en
dc.language.isoen_USen
dc.publisherAmerican Society for Microbiologyen
dc.rightsIn Copyrighten
dc.rights.holderAmerican Society for Microbiologyen
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.titleMycobacterial Interspersed Repetitive-Unit-Variable-Number Tandem-Repeat (MIRU-VNTR) Genotyping of Mycobacterium intracellulare for Strain Comparison with Establishment of a PCR-Based Databaseen
dc.title.serialJournal of Clinical Microbiologyen
dc.typeArticle - Refereeden
dc.typeDataseten
dc.type.dcmitypeTexten
dc.type.dcmitypeDataseten

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