J Korean Med Sci.  2002 Oct;17(5):593-598. 10.3346/jkms.2002.17.5.593.

Application of Infrequent-Restriction-Site Amplification for Genotyping of Mycobacterium tuberculosis and non-tuberculous Mycobacterium

• Choi TY
• Kang JO

• KMID: 1127247
• DOI: http://doi.org/10.3346/jkms.2002.17.5.593

Abstract

Infrequent restriction site amplification (IRS-PCR) is a method of amplifying DNA sequences, which flank an infrequent restriction site, and produces a strain-specific electrophoretic pattern. We studied the use of IRS-PCR to characterize Mycobacterium tuberculosis and non-tuberculous mycobactria (NTM). One-hundred and sixteen M. tuberculosis and nine NTM isolated at Hanyang University Hospital in Seoul, Korea were used in this study. IRS-PCR using AH1 and PX-G primers produced unique patterns for reference strains, M. tuberculosis H37Rv, M. bovis BCG, M. kansasii, M. scrofulaceum, M. szulgai, M. gordonae, M. avium, M. intracellulae, M. fortuitum, and M. chelonae, respectively. Reference strains M. tuberculosis H37Rv, M. bovis, M. africanum, and all isolates of M. tuberculosis showed similar IRS-PCR patterns. The IRS-PCR patterns generated with multiple isolates of M. tuberculosis from the same patients were essentially identical. IRS-PCR revealed the greatest difference between electrophoretic DNA patterns from M. avium, M. intracellulae, and M. fortuitum that differed from each other and from the reference strains. We concluded that IRS-PCR is a useful tool for strain typing of NTM, but not for M. tuberculosis.

Keyword

Mycobacterium tuberculosis; Non-tuberculous mycobacteria; Infrequent Restriction Site Amplification

MeSH Terms

Bacterial Typing Techniques/methods
Base Sequence
DNA Fingerprinting
DNA Primers/genetics
DNA, Bacterial/genetics/isolation & purification
Genotype
Humans
Korea
Mycobacterium/classification/*genetics/isolation & purification
Mycobacterium Infections/microbiology
Mycobacterium tuberculosis/classification/*genetics/isolation & purification
Polymerase Chain Reaction/*methods
Species Specificity
Tuberculosis, Pulmonary/microbiology