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Journal of Veterinary Diagnostic Investigation Vol. 18 Issue 6, 529-535
Copyright © 2006 by the American Association of Veterinary Laboratory Diagnosticians
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Full Scientific Reports

A testing scheme for the detection of Mycobacterium avium subsp. Paratuberculosis in bovine feces utilizing the ESP Para-JEM liquid culture system

Sreekumari Rajeev1, William Shulaw, Roy Berghaus, Yan Zhang and Beverly Byrum

Correspondence: 1Corresponding Author: Sreekumari Rajeev, University of Georgia, Veterinary Diagnostic and Investigational Laboratory, 43 Brighton Road, Tifton, GA

A testing scheme for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in broth cultures of bovine fecal samples carried out in ESP para-JEM System was evaluated. The scheme included acid-fast staining (on signal-positive and signal-negative samples), and confirmation by PCR for 2 MAP-specific targets and subculture of all acid-fast positive PCR-negative samples. Two hundred and fifty bovine fecal samples were evaluated for the presence of MAP using this scheme. Thirty-seven (15%) of 250 fecal samples had a positive culture result when the proposed testing scheme was used, compared to 14 (6%) positive results when using the standard ESP para-JEM protocol (requiring samples to have a positive signal from the system, a positive acid-fast stain, and a positive IS900 PCR result), and 20 (8%) positives when conventional culture was performed on Herrold egg yolk (HEY) media. A preliminary comparison of real-time and conventional PCR on DNA extracted from 15 MAP-positive broth cultures by 3 different protocols suggested that conventional PCR may be a better choice for the confirmation of the presence of MAP in the liquid cultures than real-time PCR.

Key Words: ESP • Johne disease • paratuberculosis • PCR




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Brain heart infusion broth may not be a required component for the decontamination process for the isolation of Mycobacterium avium subspecies Paratuberculosis from fecal samples using ESP(R) broth cultures
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