IS 1245 Restriction Fragment Length Polymorphism Typing of Mycobacterium avium Isolates: Proposal for Standardization

Abstract

Mycobacterium avium has become a major human pathogen, primarily due to the emergence of the AIDS epidemic. Restriction fragment length polymorphism (RFLP) typing, using insertion sequence IS1245 as a probe, provides a powerful tool in the molecular epidemiology of M. avium-related infections and will facilitate well-founded studies into the sources of M. avium infections in animal and environmental reservoirs. The standardization of this technique allows computerization of IS1245 RFLP patterns for comparison on a local level and the establishment of M. avium DNA fingerprint databases for interlaboratory comparison. Moreover, by combining international DNA typing results of M. avium complex isolates from a broad spectrum of sources, long-lasting questions on the epidemiology of this major agent of mycobacterial infections will be answered.

Molecular epidemiology of tuberculosis and other mycobacterial infections: main methodologies and achievements

“…avium [263, 269]. From the molecular epidemiological results obtained so far with the standardized IS 1245 RFLP typing [283], some conclusions can be drawn. Most serial M. avium ssp.…”

Prevalence ofMycobacterium aviumin Slaughter Pigs in The Netherlands and Comparison of IS1245Restriction Fragment Length Polymorphism Patterns of Porcine and Human Isolates

“…DNA fingerprinting. M. avium isolates were DNA fingerprinted by RFLP typing; IS1245 was used as a probe, as described previously (12,16). Internal and external molecular size markers and high-resolution gels (24 cm) were applied to facilitate computer-assisted analysis.…”

“…Computer-assisted RFLP analysis. Analysis of the IS1245 fingerprints was done with computer assistance, using GelCompar software, version 4.1 (Applied Maths, Kortrijk, Belgium), as described in a proposal for standardization of IS1245 RFLP typing (16). The band positions of the IS1245-containing restriction fragments were compared with those of a set of internal molecular weight markers by superimposing the autoradiograms of the IS1245 DNA fingerprints and the autoradiograms of the internal markers.…”

Molecular features ofMycobacterium aviumhuman isolates carrying a single copy of IS1245and IS1311per genome

“…The IS 1245 RFLP and IS 1311 RFLP analyses were performed using probes and conditions as described by van Soolingen et al (1998) and Roiz et al (1995), respectively. Probes were radiolabelled with α 32 P‐dCTP (Amersham Life Science) using the Megaprime DNA‐labelling system (Amersham Life Science).…”