Submitted to: American Society of Microbiologists Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: May 4, 1997
Publication Date: N/A
Technical Abstract: Mycobacterium paratuberculosis is the causative agent of Johne's disease, a chronic enteritis in ruminants. Currently, there is a need for improved diagnostics because of the lack of rapid and accurate identification of M. paratuberculosis-infected animals. An M. paratuberculosis genetic element was cloned, sequenced, and a species-specific oligonucleotide was synthesized. Southern hybridization analysis using the oligonucleotide identified reference, bovine, and human isolates of M. paratuberculosis. This oligonucleotide distinguished M. paratuberculosis isolates from related mycobacteria, including all M. avium strains tested in this study. The M. paratuberculosis genetic element was used in the development of a multiplex polymerase chain reaction (PCR) diagnostic test. Primers specific to the M. paratuberculosis sequence were synthesized. As an internal control to confirm that the organisms were mycobacteria, a second set of primers were synthesized based on the conserved 5' terminus of the mycobacterial RCA gene. Preliminary experiments suggest the multiplex PCR assay may be a useful tool in detection infection with mycobacteria, specifically M. paratuberculosis.