RRC ID 15833
Author Whang J, Lee BS, Choi GE, Cho SN, Kil PY, Collins MT, Shin SJ.
Title Polymerase chain reaction-restriction fragment length polymorphism of the rpoB gene for identification of Mycobacterium avium subsp. paratuberculosis and differentiation of Mycobacterium avium subspecies.
Journal Diagn. Microbiol. Infect. Dis.
Abstract Mycobacterial speciation by polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis (PRA) of the rpoB gene was evaluated for identification of Mycobacterium avium subsp. paratuberculosis (MAP) and other Mycobacterium avium complex (MAC) members to the species or subspecies level by comparison with conventional methods including hsp65 sequencing, high-performance liquid chromatography, and PCR for accepted species- or subspecies-specific genomic targets. A total of 185 type and clinical mycobacterial strains from humans, animals, and environments were tested. A 360-bp PCR product was subsequently digested with MspI, HaeIII, and SmaI restriction enzymes. The PRA using SmaI restriction showed a unique digestion pattern for MAP distinguishing it from other MAC members and other Mycobacterium spp. Moreover, HaeIII and MspI restriction of the rpoB gene enabled MAC-species and -subspecies discrimination. The rpoB-PRA using SmaI or MspI and HaeIII restriction of the rpoB gene is a simple, convenient, and reliable confirmatory assay for simultaneous identification of MAP and other MAC members.
Volume 70(1)
Pages 65-71
Published 2011-5
DOI 10.1016/j.diagmicrobio.2011.01.014
PII S0732-8893(11)00045-9
PMID 21429694
MeSH Animals Bacteriological Techniques / methods* DNA, Bacterial / genetics DNA, Bacterial / metabolism DNA-Directed RNA Polymerases / genetics* Deoxyribonuclease HpaII / metabolism Deoxyribonucleases, Type II Site-Specific / metabolism Environmental Microbiology Humans Mycobacterium avium Complex / classification* Mycobacterium avium Complex / genetics* Mycobacterium avium-intracellulare Infection / microbiology Mycobacterium avium-intracellulare Infection / veterinary Polymerase Chain Reaction / methods* Polymorphism, Restriction Fragment Length*
IF 2.341
Times Cited 5
General Microbes JCM 16228 JCM 17323 JCM 17324