Comparison of two DNA extractions and nested PCR, real-time PCR, a new commercial PCR assay, and bacterial culture for detection of Mycobacterium avium subsp. paratuberculosis in bovine feces. |
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Authors: | Jane Christopher-Hennings Matthew A Dammen Shelleen R Weeks William B Epperson Shri N Singh Gina L Steinlicht Ying Fang Jessica L Skaare Jill L Larsen Janet B Payeur Eric A Nelson |
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Affiliation: | Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, SD 57007, USA. |
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Abstract: | In this study, 5 combinations of 2 DNA extractions and 3 polymerase chain reaction (PCR) techniques were compared with culture for the detection of Mycobacterium paratuberculosis directly from bovine feces. These combinations included a new commercial extraction technique combined with a commercial PCR/Southern blot technique, nested PCR (nPCR), or real-time PCR, and a university-developed extraction combined with nPCR or real-time PCR. Four of the 5 combinations had statistically similar sensitivities between 93% and 100% and specificity between 95% and 100%, when compared with culture results from 63 bovine fecal samples. These results indicated that using a commercial extraction with a commercial PCR/Southern blot, nPCR, or real-time PCR, or a university-developed extraction with real-time PCR would result in similar sensitivities to culture for the identification of M. paratuberculosis from bovine feces and are valid alternatives to culture. |
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