Development of a diagnostic test for Johne's disease using a DNA hybridization probe

Abstract

A DNA probe, M13 mpHAW71, that detects Mycobacterium paratuberculosis in the fecal material of infected animals was developed for use in the diagnosis of Johne's disease. The probe detected as few as IO' M. paratuberculosis when hybridized under stringent conditions to total genomic DNA purified from bovine fecal material. When the probe was used diagnostically, it did not differentiate members of the Mycobacterium avium-M. intracellulare-M. paratuberculosis complex. Compared with culturing, the DNA probe identified 34.4% more mycobacterium-containing fecal samples, and testing took only 72 h to complete.

Comparison of immunomagnetic bead separation‐polymerase chain reaction and faecal culture for the detection of Mycobacterium avium subsp paratuberculosis in sheep faeces

“…Earlier attempts to detect M a paratuberculosis directly in faecal samples by PCR have had limited success, limited application or were not reproducible. [17][18][19][20] Although most of and decrease milk production. 1 Aborted bovine foetuses show nonspecific lesions and various degrees of autolysis.…”

How does a Mycobacterium change its spots? Applying molecular tools to track diverse strains of Mycobacterium avium subspecies paratuberculosis

“…All members of the MAC are genetically similar, although the small distinctions between Mycobacterium avium and Mycobacterium intracellulare have been well established (Boddinghaus et al 1990;De Smet et al 1996;Feizabadi et al 1997;Thorel et al 2001). In particular, DNA-DNA hybridization studies have long ago showed significant genetic similarity between MAP and MAA (Saxegaard et al 1988; Yoshimura and Graham 1988;Hurley et al 1989). More recently, a >98% nucleotide identity was shown to exist when comparing MAP and MAA genomes with each other .…”

Detection of Mycobacterium Avium Subsp. Paratuberculosis in Bovine Fecal Samples: Comparison of Three Polymerase Chain Reaction—Based Diagnostic Tests with a Conventional Culture Method

“…Several DNA probes for the detection of MAP have been described. 13,19,21 More recently, several polymerase chain reaction (PCR) tests to detect MAP DNA have been developed. 1,5,8,23,27,29 The purpose of this study was to compare 3 PCR-based diagnostic kits with a conventional culture method for the detection of MAP in bovine fecal samples.…”