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Molecular tools for the characterization of mycobacterium avium subspecies paratuberculosis

dc.contributor.committeeMemberWoodbury, Murray R.en_US
dc.contributor.committeeMemberPolley, Lyddenen_US
dc.contributor.committeeMemberDeneer, Harryen_US
dc.contributor.committeeMemberChirino-Trejo, Manuelen_US
dc.contributor.committeeMemberAppleyard, Greg D.en_US
dc.creatorSibley, Jennifer Anneen_US
dc.date.accessioned2005-03-29T00:09:22Zen_US
dc.date.accessioned2013-01-04T04:27:29Z
dc.date.available2005-04-04T08:00:00Zen_US
dc.date.available2013-01-04T04:27:29Z
dc.date.created2005-01en_US
dc.date.issued2005-01-28en_US
dc.date.submittedJanuary 2005en_US
dc.description.abstractSeveral strain typing techniques are available to categorize Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) isolates into cattle, sheep, bison, and Intermediate groups. The majority of isolates studied were identified as members of the cattle associated group, regardless of sample host origin, suggesting that the cattle group of M. paratuberculosis isolates are very successful. This may be because host specificity is not critical for this group or the small differences required to demonstrate host specificity have not yet been found. A major limitation to the epidemiological study of M. paratuberculosis has been the difficulty associated with laboratory cultivation of this micro-organism. The new typing techniques described in this thesis do not require viable M. paratuberculosis bacteria and therefore open a door to novel typing practices. The new molecular techniques, single stranded conformation polymorphism (SSCP) analysis and satellite typing, were applied to M. paratuberculosis isolates (n=75) from a broad range of ruminant hosts and geographic locations. SSCP analysis and satellite typing were compared to currently accepted techniques (PCR-REA, RFLP, PFGE) for their ability to rapidly and reliably differentiate among M. paratuberculosis isolates. PCR-REA segregated isolates (n=75) into cattle (n=72), sheep (n=1) or bison (n=2) associated strain types. Two isolates from cattle in Canada were typed as RFLP-BstEII C5 by RFLP analysis. PFGE grouped a subset (n=8) of M. paratuberculosis isolates into 4 different PFGE types. Satellite typing resulted in 4 different satellite types (A, B, C, D). SSCP analysis identified 2 regions (IS900-2 and HSP70) where sequence polymorphisms could be targeted to display differences among M. paratuberculosis isolates.en_US
dc.identifier.urihttp://hdl.handle.net/10388/etd-03292005-000922en_US
dc.language.isoen_USen_US
dc.subjectMicrosatellitesen_US
dc.subjectJohnes Diseaseen_US
dc.subjectMolecularen_US
dc.subjectPCRen_US
dc.titleMolecular tools for the characterization of mycobacterium avium subspecies paratuberculosisen_US
dc.type.genreThesisen_US
dc.type.materialtexten_US
thesis.degree.departmentVeterinary Microbiologyen_US
thesis.degree.disciplineVeterinary Microbiologyen_US
thesis.degree.grantorUniversity of Saskatchewanen_US
thesis.degree.levelMastersen_US
thesis.degree.nameMaster of Science (M.Sc.)en_US

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