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Differentiation of Mycobacterial species by hsp65 duplex PCR followed by duplex-PCR-based restriction analysis and direct sequencing

Cited 17 time in Web of Science Cited 18 time in Scopus
Authors
Kim, Hyun-Ju; Mun, Ho-Sok; Kim, Hong; Oh, Eun-Ju; Ha, Youngju; Bai, Gill-Han; Park, Young-Gil; Cha, Chang-Yong; Kook, Yoon-Hoh; Kim, Bum-Joon
Issue Date
2006-08-25
Publisher
American Society for Microbiology
Citation
J Clin Microbiol. 2006 Nov;44(11):3855-62. Epub 2006 Aug 23.
Keywords
Bacterial Proteins/*geneticsChaperonins/*geneticsHumansMycobacterium tuberculosis/*classification/geneticsPolymerase Chain Reaction/*methodsSensitivity and Specificity*Sequence Analysis, DNA
Abstract
Here we describe a novel duplex PCR method which can differentiate Mycobacterium tuberculosis and nontuberculosis mycobacteria (NTM) strains by amplifying hsp65 DNAs of different sizes (195 and 515 bp, respectively). The devised technique was applied to 54 reference and 170 clinical isolates and differentiated all strains into their respective groups with 100% sensitivity and specificity. Furthermore, a duplex PCR-restriction analysis (duplex PRA) and a direct sequencing protocol were developed to differentiate NTM strains at the species and subspecies levels based on previously reported hsp65 DNA sequences (H. Kim et al., Int. J. Syst. Evol. Microbiol. 55:1649-1656, 2005) and then applied to 105 NTM clinical isolates. All NTM isolates were clearly differentiated at the species and subspecies levels by subsequent procedures (PRA or direct sequencing) targeting 515-bp NTM duplex PCR amplicons. Our results suggest that novel duplex PCR-based methods are sensitive and specific for identifying mycobacterial culture isolates at the species level.
ISSN
0095-1137 (Print)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16928964

http://hdl.handle.net/10371/29677
DOI
https://doi.org/10.1128/JCM.01214-06
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College of Medicine/School of Medicine (의과대학/대학원)Microbiology (미생물학전공)Journal Papers (저널논문_미생물학전공)
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