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Clinical and Vaccine Immunology, September 2009, p. 1314-1321, Vol. 16, No. 9
1071-412X/09/$08.00+0     doi:10.1128/CVI.00151-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Assessment of Mycobacterium tuberculosis OmpATb as a Novel Antigen for the Diagnosis of Bovine Tuberculosis{triangledown}

Irene Schiller,1 H. Martin Vordermeier,2 W. Ray Waters,3 Mitchell Palmer,3 Tyler Thacker,3 Adam Whelan,2 Roland Hardegger,1 Beatrice Marg-Haufe,1 Alex Raeber,1 and Bruno Oesch1*

Prionics AG, Schlieren, Switzerland,1 Veterinary Laboratory Agency, Addlestone, Great Britain,2 National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, Iowa3

Received 3 April 2009/ Returned for modification 19 May 2009/ Accepted 1 July 2009

In the search for better tools to control bovine tuberculosis, the development of diagnostic tests with improved specificity and sensitivity has a high priority. We chose to search for novel immunodiagnostic reagents. In this study, Rv0899 (outer membrane protein A of Mycobacterium tuberculosis [OmpATb]) was evaluated as a stimulation antigen in a gamma interferon (IFN-{gamma}) release assay to diagnose bovine tuberculosis. OmpATb induced IFN-{gamma} responses in cattle experimentally infected with M. bovis as early and as persistently as ESAT-6 and CFP-10, the current lead diagnostic antigens. In naturally infected cattle, OmpATb stimulated IFN-{gamma} production in 22 of 26 animals (85%). Importantly, OmpATb detected a portion of M. bovis-infected cattle which did not respond to ESAT-6 and CFP-10 (five of six cattle). The combined diagnostic sensitivity of OmpATb, ESAT-6, and CFP-10 for a preselected group consisting of naturally infected cattle with an overrepresentation of ESAT-6/CFP-10 nonresponders was 96% (25 of 26 animals). The specificity of OmpATb for uninfected cattle was 100% (27 cattle were tested; 12 of them gave false-positive results with tuberculins). In summary, our results indicate that OmpATb has the potential to enhance the sensitivity of previously described diagnostic tests based on ESAT-6 and CFP-10 and that the combined use of OmpATb, ESAT-6, CFP-10, and other proteins may achieve at least equal sensitivity to that obtained with purified protein derivative, but at a higher specificity. Further studies evaluating the diagnostic performance of OmpATb in combination with other proteins are ongoing.

* Corresponding author. Mailing address: Prionics AG, Wagistrasse 27A, CH-8952 Schlieren, Switzerland. Phone: 41 44 200 2014. Fax: 41 44 200 2010. E-mail: bruno.oesch{at}prionics.com

{triangledown} Published ahead of print on 8 July 2009.

Clinical and Vaccine Immunology, September 2009, p. 1314-1321, Vol. 16, No. 9
1071-412X/09/$08.00+0     doi:10.1128/CVI.00151-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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