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 |
2011-5-1
|
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 |
2.499
|
Times Cited |
5
|
WOS Category
|
INFECTIOUS DISEASES
MICROBIOLOGY
|
Resource |
General Microbes |
JCM 16228
JCM 17323
JCM 17324 |