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Possibilities for improving the D-PCR test for M. a. paratuberculosis from faeces
The rapid diagnosis of M. a. paratuberculosis infection in the early stages of disease, or where prevalence is low, is still critical for the management of ovine Johne's disease. Recently we reported the ability to detect M. a. paratuberculosis directly from sheep faecal samples by PCR (D-PCR) but the sensitivity of this test was less than the current gold standard test, pooled faecal culture (faecal culture). This project was undertaken to improve the sensitivity of the D-PCR test so that it was greater than or equal to faecal culture, without increasing the cost to the producer. Unfortunately, the new D-PCR described in this report failed to meet this objective. However, another D-PCR test developed in Japan that utilizes real-time PCR technology was also evaluated in the concluding stages of the project. The preliminary results with this test suggest that it has sensitivity equivalent to or greater than faecal culture. We highly recommended that this D-PCR test be further examined and developed for Australian conditions as a replacement or substitute test for faecal culture. As part of this project we successfully developed a new conventional multiplex PCR for M. a. paratuberculosis that includes an internal control. This new conventional PCR also warrants further investigation and should also be included in the future development of the D-PCR test to provide improved diagnosis and quality assurance for laboratories that are currently unable to use real-time-PCR technology for routine diagnostics.
This page was last updated on 24/07/2017
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