Essay
Characterization and development of an assay for mycobacterium paratuberculosis
2002
Handle:
https://hdl.handle.net/2376/2400
Abstract
Mycobacterium avium subspecies paratuberculosis, the causative agent of Johne's disease, is a bacterium of significant medical and economic importance to cattle and sheep producers. In contrast to cattle, diagnosis of sub-clinical and clinical ovine M. paratuberculosis infections is extremely difficult. In sheep, the clinical signs are vague, as a general loss of body condition often occurs unaccompanied by the characteristic diarrheal symptoms of cattle. Current serological tests implemented in bovine diagnosis are often inaccurate in detecting ovine strains. Moreover, ovine strains of M. paratuberculosis do not grow well, or at all, in culture.
To date, the reason for these differences is unknown. Nevertheless, the phenotypic differences are most likely linked to genotypic variation. The aim of this project was to evaluate a well characterized and distinct genetic marker of the mycobacterial genome, the 65kDa heat shock protein gene (hsp65) , for sequence variations between bovine and ovine strains. Sequence variations have the potential to be used in the development of a polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) based assay that can detect and differentiate ovine strains of M. paratuberculosis. Ovine M. paratuberculosis isolates were obtained from the National Animal Disease Center (Ames, Iowa) and the RFLP profile of the ovine hsp65 gene compared with those of bovine isolates. The ovine hsp65 gene was sequenced and the sequence analyzed for novel restriction endonuclease cut sites. One enzyme candidate, Faul, was chosen for digestion of the ovine and bovine samples. Analysis of the RFLP revealed discemable differences between the ovine and bovine sequences that could be used as the basis for a combination PCR and RFLP based assay for ovine M. paratuberculosis.
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Details
- Title
- Characterization and development of an assay for mycobacterium paratuberculosis
- Creators
- Marlene K. Bakko (Author)
- Contributors
- Inge Eriks (Advisor)
- Academic Unit
- Honors Theses (WSU Pullman, Passed with Distinction)
- Identifiers
- 99900590747401842
- Copyright
- http://rightsstatements.org/vocab/InC/1.0/; http://www.ndltd.org/standards/metadata; http://purl.org/eprint/accessRights/OpenAccess; In copyright; Publicly accessible; openAccess
- Language
- English
- Resource Type
- Essay