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PCR restriction fragment length polymorphism analysis (PRA)-algorithm targeting 644 bp Heat Shock Protein 65 (hsp65) gene for differentiation of Mycobacterium spp

Cited 34 time in Web of Science Cited 35 time in Scopus
Issue Date
2005-07-13
Publisher
Elsevier
Citation
J Microbiol Methods. 2005 Aug;62(2):199-209.
Keywords
AlgorithmsBacterial Proteins/chemistry/*geneticsChaperonins/chemistry/*geneticsDNA, Bacterial/chemistry/geneticsHumansMycobacterium/*classification/genetics/isolation & purificationMycobacterium Infections/microbiologyPolymerase Chain Reaction/*methodsPolymorphism, Restriction Fragment LengthSequence Analysis, DNA
Abstract
A method based on PCR-restriction fragment length polymorphism analysis (PRA) using a novel region of the hsp65 gene was developed for the rapid and exact identification of mycobacteria to the species level. A 644 bp region of hsp65 in 62 mycobacteria reference strains, and 4 related bacterial strains were amplified, and the amplified DNAs were subsequently digested with restriction enzymes, namely, AvaII, HphI, and HpaII. Most of the mycobacteria species were easily differentiated at the species level by the developed method. In particular, the method enabled the separation of M. avium, M. intracellulare and M. tuberculosis to the species level by AvaII digestion alone. An algorithm was constructed based on the results and a blind test was successfully performed on 251 clinical isolates, which had been characterized by conventional biochemical testing. Our results suggest that this novel PRA offers a simple, rapid, and accurate method for the identification of mycobacteria culture isolates at the species level.
ISSN
0167-7012 (Print)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16009277

https://hdl.handle.net/10371/23424
DOI
https://doi.org/10.1016/j.mimet.2005.02.010
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College of Medicine/School of Medicine (의과대학/대학원)Microbiology (미생물학전공)Journal Papers (저널논문_미생물학전공)
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