An extensive field comparison of the gamma interferon (IFN-gamma) assay and the single intradermal tuberculin test for the diagnosis of bovine tuberculosis was conducted in Australia. The specificity of the IFN-gamma assay was determined by testing more than 6000 cattle from tuberculosis-free herds and varied from 96.2% to 98.1%, depending on the cut-off point chosen to define a positive reactor. For the sensitivity trial, cattle from herds being de-populated because of bovine tuberculosis were examined with both assays. The sensitivity of the IFN-gamma assay was shown to be significantly higher than the single intradermal tuberculin test and varied from 76.8% to 93.6% depending on the method of interpretation. A maximum overall sensitivity of 95.2% was obtained by testing with the IFN-gamma and the tuberculin test in parallel. The superior sensitivity of the IFN-gamma assay and the ability to adjust the sensitivity of the system depending on the task involved, will provide the Australian Tuberculosis Eradication Campaign with a valuable additional test to enable it to accomplish its goals.
A DNA amplification assay using the polymerase chain reaction technique designed for the rapid identification of Mycobacterium bovis organisms was used to test 211 human mycobacterial isolates and 177 clinical specimens previously submitted for routine mycobacterial culture. The procedures described could be used by routine or specialist laboratories for identification of M. tuberculosis complex organisms in 4 h and/or as a rapid screening method for the direct detection of M. tuberculosis complex organisms in specimens.
The BACTEC radiometric culture method for detection of Mycobacterium paratuberculosis was evaluated on faeces from cattle on a farm in quarantine for Johne's disease. A multiplex polymerase chain reaction (PCR) based on the IS900 sequence specific for M paratuberculosis and a genus specific 16S rRNA region was developed and used to test cultures showing evidence of mycobacterial growth in the BACTEC liquid radiometric culture medium. Using the BACTEC-PCR combination, confirmation of M paratuberculosis from faeces and tissue of clinically affected animals was achieved within 2 to 4 weeks and 1 week, respectively, a substantial improvement on traditional culture and identification methods. The PCR provided rapid exclusion of M paratuberculosis when other Mycobacterium spp were grown. The radiometric culture medium proved to be very sensitive for culturing Mycobacterium spp.
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