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Clinical and Vaccine Immunology, August 2008, p. 1277-1281, Vol. 15, No. 8
1071-412X/08/$08.00+0     doi:10.1128/CVI.00105-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Diagnosis of Bovine Paratuberculosis by a Novel Enzyme-Linked Immunosorbent Assay Based on Early Secreted Antigens of Mycobacterium avium subsp. paratuberculosis{triangledown}

Sung Jae Shin,1,2 Donghee Cho,2,3 and Michael T. Collins2*

Department of Microbiology and Infection Signaling Network Research Center, College of Medicine, Chungnam National University, Daejeon 301-747, South Korea,1 Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin—Madison, 2015 Linden Drive, Madison, Wisconsin 53706-1102,2 Bacteriology & Parasitology Division, Animal Disease Research Department, National Veterinary Research and Quarantine Service, Anyang, Kyounggi 430-824, South Korea3

Received 25 March 2008/ Returned for modification 29 April 2008/ Accepted 30 May 2008

We previously reported that protein antigens of serodiagnostic potential were more abundant in culture filtrates than cellular extracts from liquid cultures of Mycobacterium avium subsp. paratuberculosis (D. Cho and M. T. Collins, Clin. Vaccine Immunol. 13:1155-1161, 2006). Based on this observation, a novel enzyme-linked immunosorbent assay (ELISA) using antigens secreted by young (early- to mid-log-phase) cultures of M. avium subsp. paratuberculosis JTC303 (a low-passage isolate originating from the ileum of a Holstein bull) in mycobactin-supplemented Watson-Reid medium (pH 6.0) was developed and evaluated using a previously described panel of bovine sera (M. T. Collins et al., Clin. Diagn. Lab. Immunol. 12:685-692, 2005) that included 444 paratuberculosis cases and 412 controls. The new assay, called JTC-ELISA, had a significantly higher diagnostic sensitivity and an equivalent specificity compared to those of five commercial paratuberculosis ELISA kits. By receiver-operating characteristic analysis, the JTC-ELISA had the highest area under the curve of the six assays evaluated. The JTC-ELISA was particularly sensitive at detecting low-level fecal shedders of Mavium subsp. paratuberculosis (40%; the sensitivity of the commercial kits was 20%). The JTC-ELISA works effectively on both serum and milk samples for the detection of cattle with subclinical M. avium subsp. paratuberculosis infections, providing a cost-effective diagnostic tool to support paratuberculosis control programs in cattle herds.

* Corresponding author. Mailing address: 2015 Linden Drive, Madison, WI 53706-1102. Phone: (608) 262-8457. Fax: (608) 265-6463. E-mail: mcollin5{at}wisc.edu

{triangledown} Published ahead of print on 11 June 2008.

Clinical and Vaccine Immunology, August 2008, p. 1277-1281, Vol. 15, No. 8
1071-412X/08/$08.00+0     doi:10.1128/CVI.00105-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Shin, S. J., Anklam, K., Manning, E. J. B., Collins, M. T. (2009). Rapid Mycobacterial Liquid Culture-Screening Method for Mycobacterium avium Complex Based on Secreted Antigen-Capture Enzyme-Linked Immunosorbent Assay. CVI 16: 613-620 [Abstract] [Full Text]  


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