CVI
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zheng, L.
Right arrow Articles by Minocha, H. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zheng, L.
Right arrow Articles by Minocha, H. C.

 Previous Article  |  Next Article 

Clinical and Diagnostic Laboratory Immunology, July 2000, p. 557-562, Vol. 7, No. 4
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Cloning of the Bovine Immunodeficiency Virus gag Gene and Development of a Recombinant-Protein-Based Enzyme-Linked Immunosorbent Assay†

Ling Zheng,1 Michelle Swanson,1 Jinghua Liao,1 Charles Wood,2 Sanjay Kapil,1 Ron Snider,3 Thomas A. Loughin,4 and Harish C. Minocha1,*

Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine,1 and Department of Statistics, College of Arts and Sciences,4 Kansas State University, Manhattan, Kansas 66506; School of Biological Sciences, University of Nebraska, Lincoln, Nebraska 685882; and Department of Veterinary Pathology, School of Veterinary Medicine, and Department of Dairy Science, Louisiana State University, Baton Rouge, Louisiana 708033

Received 28 December 1999/Returned for modification 8 February 2000/Accepted 24 March 2000

An enzyme-linked immunosorbent assay (ELISA) was established for the rapid detection of specific bovine immunodeficiency virus (BIV) antibodies in cattle, using recombinant Gag protein as an antigen. The gag coding region from BIV was cloned into an expression vector, pQE32, which expressed high levels of recombinant protein from Escherichia coli. The ELISA was standardized by a checkerboard titration against known BIV-positive and -negative sera from cattle and a monoclonal antibody to the Gag protein. A total of 139 cattle serum samples, from the diagnostic laboratory at Kansas State University, Manhattan, and from the Dairy Station, Louisiana State University, Baton Rouge, were compared by ELISA and immunoblot assays for the detection of BIV-specific antibodies. Of 26 cattle sera samples which tested positive using the immunoblot assay, 23 were positive by ELISA, thus establishing a strong correlation between the two tests. The sensitivity and specificity of ELISA relative to immunoblotting were 0.88 and 0.93, respectively. ELISA proved to be as specific as immunoblotting but was much less time-consuming and easier to perform.

* Corresponding author. Mailing address: Department of Diagnostic Medicine-Pathobiology, College of Veterinary Medicine, 1800 Denison Ave., Kansas State University, Manhattan, KS 66506. Phone: (785) 532-4603. Fax: (785) 532-4039. E-mail: Minocha{at}vet.ksu.edu.

dagger Contribution 99-55-J from the Kansas Agriculture Experiment Station.

Clinical and Diagnostic Laboratory Immunology, July 2000, p. 557-562, Vol. 7, No. 4
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Antimicrob. Agents Chemother. Clin. Microbiol. Rev. Infect. Immun.
J. Clin. Microbiol. J. Virol. ALL ASM JOURNALS

Copyright © 2000 by the American Society for Microbiology. All rights reserved.