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dc.contributor.authorSubedi, Shradha
dc.contributor.authorKong, Fanrong
dc.contributor.authorJelfs, Peter
dc.contributor.authorGray, Timothy J
dc.contributor.authorXiao, Meng
dc.contributor.authorSintchenko, Vitali
dc.contributor.authorChen, Sharon C-A
dc.date.accessioned2020-02-20T03:32:14Z
dc.date.available2020-02-20T03:32:14Z
dc.date.issued2016
dc.identifier.issn1932-6203
dc.identifier.doi10.1371/journal.pone.0164138
dc.identifier.urihttp://hdl.handle.net/10072/391715
dc.description.abstractAccurate identification of slowly growing nontuberculous mycobacteria (SG-NTM) of clinical significance remains problematic. This study evaluated a novel method of SG-NTM identification by amplification of the mycobacterial 16S-23S rRNA internal transcribed spacer (ITS) region followed by resolution of amplified fragments by sequencer-based capillary gel electrophoresis (SCGE). Fourteen American Type Culture Collection (ATCC) strains and 103 clinical/environmental isolates (total n = 24 species) of SG-NTM were included. Identification was compared with that achieved by high performance liquid chromatography (HPLC), in-house PCR and 16S/ITS sequencing. Isolates of all species yielded a SCGE profile comprising a single fragment length (or peak) except for M. scrofulaceum (two peaks). SCGE peaks of ATCC strains were distinct except for peak overlap between Mycobacterium kansasii and M. marinum. Of clinical/environmental strains, unique peaks were seen for 7/17 (41%) species (M. haemophilum, M. kubicae, M. lentiflavum, M. terrae, M. kansasii, M. asiaticum and M. triplex); 3/17 (18%) species were identified by HPLC. There were five SCGE fragment length types (I–V) each of M. avium, M. intracellulare and M. gordonae. Overlap of fragment lengths was seen between M. marinum and M. ulcerans; for M. gordonae SCGE type III and M. paragordonae; M. avium SCGE types III and IV, and M. intracellulare SCGE type I; M. chimaera, M. parascrofulaceum and M. intracellulare SCGE types III and IV; M. branderi and M. avium type V; and M. vulneris and M. intracellulare type V. The ITS-SCGE method was able to provide the first line rapid and reproducible species identification/screening of SG-NTM and was more discriminatory than HPLC.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherPublic Library of Science
dc.relation.ispartofissue10
dc.relation.ispartofjournalPLoS One
dc.relation.ispartofvolume11
dc.subject.keywordsScience & Technology
dc.subject.keywordsMultidisciplinary Sciences
dc.subject.keywordsScience & Technology - Other Topics
dc.subject.keywordsRIBOSOMAL-RNA
dc.subject.keywordsAVIUM COMPLEX
dc.title16S-23S Internal Transcribed Spacer Region PCR and Sequencer-Based Capillary Gel Electrophoresis has Potential as an Alternative to High Performance Liquid Chromatography for Identification of Slowly Growing Nontuberculous Mycobacteria
dc.typeJournal article
dc.type.descriptionC1 - Articles
dcterms.bibliographicCitationSubedi, S; Kong, F; Jelfs, P; Gray, TJ; Xiao, M; Sintchenko, V; Chen, SC-A, 16S-23S Internal Transcribed Spacer Region PCR and Sequencer-Based Capillary Gel Electrophoresis has Potential as an Alternative to High Performance Liquid Chromatography for Identification of Slowly Growing Nontuberculous Mycobacteria, PLoS One, 2016, 11 (10)
dcterms.dateAccepted2016-09-20
dcterms.licensehttp://creativecommons.org/licenses/by/4.0/
dc.date.updated2020-02-20T03:30:07Z
dc.description.versionVersion of Record (VoR)
gro.rights.copyright© 2016 Subedi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
gro.hasfulltextFull Text
gro.griffith.authorSubedi, Shradha


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