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Journal of Clinical Microbiology, April 2008, p. 1207-1212, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.02031-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Sequence Polymorphisms in a Surface PPE Protein Distinguish Types I, II, and III of Mycobacterium avium subsp. paratuberculosis

Department of Medicine, Division of Gastroenterology, Faculty of Medicine, University of Calgary, Calgary, Alberta T2N 4N1, Canada,¹ Department of Production Animal Health, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta T2N 4N1, Canada²

Received 17 October 2007/ Returned for modification 23 December 2007/ Accepted 4 February 2008

In the last 2 decades, a variety of different molecular typing methods have been developed to differentiate strains of Mycobacterium avium subsp. paratuberculosis. The most successful techniques are based on insertion sequences, repetitive loci, comparative genomics, or single nucleotide polymorphisms. In the present study, we chose to examine whether a single M. avium subsp. paratuberculosis gene could serve as a means of differentiation of a variety of isolates. The MAP1506 gene locus encodes a member of the polymorphic PPE protein family that has putative roles relevant to M. avium subsp. paratuberculosis pathogenicity. The MAP1506 locus was sequenced from a collection of 58 M. avium subsp. paratuberculosis isolates from different sources, hosts, and typing profiles. Following sequence alignment and analysis, it was found that bovine (type II) strains of M. avium subsp. paratuberculosis consistently differed from ovine (type I) and intermediate (type III) strains in seven and eight nucleotides, respectively. Polymorphic regions of the MAP1506 locus were selected for analysis by denaturing gradient gel electrophoresis, allowing visual discrimination of the three subtypes of M. avium subsp. paratuberculosis isolates. This is the first report describing the use of PCR and denaturing gradient gel electrophoresis on a single gene as a method to distinguish types I, II, and III of M. avium subsp. paratuberculosis.

Published ahead of print on 13 February 2008.