Title

Characterization Of A Specific Mycobacterium Paratuberculosis Recombinant Clone Expressing 35,000-Molecular-Weight Antigen And Reactivity With Sera From Animals With Clinical And Subclinical Johne'S Disease

Authors

Authors

F. A. K. ElZaatari; S. A. Naser;D. Y. Graham

Comments

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Abbreviated Journal Title

J. Clin. Microbiol.

Keywords

POLYMERASE CHAIN-REACTION; CROSSED IMMUNOELECTROPHORETIC ANALYSIS; LINKED-IMMUNOSORBENT-ASSAY; COMMERCIAL DNA PROBE; BOVINE; PARATUBERCULOSIS; CROHNS-DISEASE; PARA-TUBERCULOSIS; ESCHERICHIA-COLI; CONTROL TISSUES; DIAGNOSIS; Microbiology

Abstract

Johne's disease is a chronic enteritis of ruminants associated with enormous worldwide economic losses for the dairy cow- and goat-rearing industries, Management limitations and eradication programs for this disease have been hampered by the lack of a simple and specific diagnostic test for the detection of subclinical cases, We used a recombinant clone expressing a 35,000-molecular-weight Mycobacterium paratuberculosis antigen (p35 antigen) from a previously constructed expression library of M. paratuberculosis in Escherichia coli, The DNA fragment encoding the p35 gene hybridized only to DNA from Mycobacterium avium complex, but not to DNAs from other mycobacteria and nonmycobacterial organisms, The seroreactivity of p35 was evaluated by immunoblotting against 57 reference serum samples obtained from infected and uninfected animals, p35 was recognized by sera from 100% of animals with advanced Johne's disease (clinical stage) (12 cattle, 2 goats, and 2 sheep) and by sera from 75% of 20 cattle with early infection (subclinical stage), None of the sera from 15 M. paratuberculosis-free cows, 3 Mycobacterium bovis BCG-infected tuberculous cattle, or 3 cows artificially inoculated with multiple doses of viable M. paratuberculosis reacted with p35. The overall sensitivity, specificity, positive predictive value, and negative predictive value were 86, 100, 100, and 75%, respectively. The accuracy of p35 immunoblotting was superior to those of commercially available diagnostic tests for Johne's disease. These results suggest that the p35 recombinant protein has potential for use in the serodiagnosis of animals with Johne's disease at all stages of infection, The DNA fragment encoding p35 may also serve as a probe for identification of M. avium complex infection.

Journal Title

Journal of Clinical Microbiology

Volume

35

Issue/Number

7

Publication Date

1-1-1997

Document Type

Article

Language

English

First Page

1794

Last Page

1799

WOS Identifier

WOS:A1997XE59100030

ISSN

0095-1137

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