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Typing of Mycobacterium avium Subspecies paratuberculosis Isolates from Newfoundland Using Fragment Analysis
Authors:Milka P. Podder  Susan E. Banfield  Greg P. Keefe  Hugh G. Whitney  Kapil Tahlan
Affiliation:1. Department of Biology, Memorial University of Newfoundland, St. John’s, Newfoundland and Labrador, Canada.; 2. Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, Prince Edward Island, Canada.; 3. Animal Health Division, Newfoundland and Labrador Department of Natural Resources, St. John''s, Newfoundland and Labrador, Canada.; St. Petersburg Pasteur Institute, RUSSIAN FEDERATION,
Abstract:Short Sequence Repeat (SSR) typing of Mycobacterium avium subspecies paratuberculosis (Map) isolates is one of the most commonly used method for genotyping this pathogen. Currently used techniques have challenges in analyzing mononucleotide repeats >15 bp, which include some of the Map SSRs. Fragment analysis is a relatively simple technique, which can accurately measure the size of DNA fragments and can be used to calculate the repeat length of the target SSR loci. In the present study, fragment analysis was used to analyze 4 Map SSR loci known to provide sufficient discriminatory power to determine the relationship between Map isolates. Eighty-five Map isolates from 18 animals from the island of Newfoundland were successfully genotyped using fragment analysis. To the best of our knowledge, this is the first report on Map SSR diversity from Newfoundland dairy farms. Previously unreported Map SSR-types or combinations were also identified during the course of the described work. In addition, multiple Map SSR-types were isolated from a single animal in many cases, which is not a common finding.
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