Polymerase chain reaction-restriction fragment length polymorphism of the rpoB gene for identification of Mycobacterium avium subsp. paratuberculosis and differentiation of Mycobacterium avium subspecies

Jake Whang, Byung Soo Lee, Go Eun Choi, Sang Nae Cho, Park Young Kil, Michael T. Collins, Sung Jae Shin

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)65-71
Number of pages7
JournalDiagnostic Microbiology and Infectious Disease
Volume70
Issue number1
DOIs
Publication statusPublished - 2011 May

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)
  • Infectious Diseases

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