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

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

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)87-93
Number of pages7
JournalJournal of Veterinary Diagnostic Investigation
Volume15
Issue number2
DOIs
StatePublished - Mar 2003
Externally publishedYes

ASJC Scopus subject areas

  • veterinary(all)

Fingerprint Dive into the research topics of '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'. Together they form a unique fingerprint.

Cite this