Use of Multilocus Variable-Number Tandem-Repeat Analysis for TypingMycobacterium aviumsubsp.paratuberculosis

Abstract

The etiology of Crohn's disease in humans is largely unknown. Clinical signs of Crohn's disease partly resemble the clinical picture of Johne's disease in ruminants caused by Mycobacterium avium subsp. paratuberculosis. Because of the high prevalence of these bacteria in (products of) ruminants and their remarkable thermostability, concern has been raised about the possible role of these bacteria in the pathogenesis of Crohn's disease. In an attempt to develop a molecular typing method to facilitate meaningful comparative DNA fingerprinting of M. avium subsp. paratuberculosis isolates from the human and animal reservoirs, multilocus variable-number tandem-repeat analysis (MLVA) was explored and compared to IS900 restriction fragment length polymorphism (RFLP) typing. MLVA typing subdivided the most predominant RFLP type, R01, into six subtypes and thus provides a promising molecular subtyping approach to study the diversity of M. avium subsp. paratuberculosis

Prevalence of avian influenza viruses,Borrelia garinii,Mycobacterium avium, andMycobacterium aviumsubsp.paratuberculosisin waterfowl and terrestrial birds in Slovakia, 2006

“…Primers specific for Mycobacteria were described previously by Telenti et al (1993). Birds were screened for MAP using primers IS900A and IS900B previously described by Overduin et al (2004). All specific PCR products were separated by electrophoresis through 2% agarose gels and were visualized by staining with ethidium bromide.…”

Identification and Characterization of Variable-Number Tandem-Repeat Markers for Typing of Brucella spp

“…As a result, this approach has proven particularly useful as a tool for strain discrimination in bacterial species with little genomic variation, notably Bacillus anthracis (17), Yersinia pestis (17), Francisella tularensis (16), and various mycobacterial species such as Mycobacterium tuberculosis (22), Mycobacterium leprae (15), and Mycobacterium avium subsp. paratuberculosis (20). As well as the potential for high resolution, MLVA has a number of other technical advantages, most notably its relative simplicity, fast turnaround time, amenability to high-throughput approaches, applicability to nonviable and/or crude preparations, and the ability to easily compare digital results between laboratories.…”

Use of an Automated Multiple-Locus, Variable-Number Tandem Repeat-Based Method for Rapid and High-Throughput Genotyping of Staphylococcus aureus Isolates

“…The discriminatory power of our eight target-based VNTR approach appears to be appreciable, as groups of strains sharing identical SmaI PFGE profiles appeared to be only related and not clonal, and they remained segregated in subdivisions of large clusters. The simplicity of this method, which relies on a single multiplex PCR amplification and a totally automated clustering analysis, limits the risk of user-dependent variation, such as that encountered in image analysis-based techniques (3,26). As suggested recently (27), PCR-based techniques dedicated to genotyping are powerful and allow identifications to be made rapidly, even when slowly growing bacteria are studied.…”

“…Direct digestion of the intact chromosome followed by size separation (pulsed-field gel electrophoresis [PFGE]) or genotyping by nucleic acid probes (ribotyping) are two techniques that do not require PCR amplification. However, PCR-based techniques appear to be advantageous for the study of slowly growing or difficult-to-grow organisms (26).…”