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Journal of Clinical Microbiology, November 2000, p. 4010-4014, Vol. 38, No. 11
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development of a Highly Sensitive and Specific Enzyme-Linked Immunosorbent Assay Based on Recombinant Matrix Protein for Detection of Avian Pneumovirus Antibodies

Baldev R. Gulati,1 Kjerstin T. Cameron,2 Bruce S. Seal,3 Sagar M. Goyal,1 David A. Halvorson,2 and M. Kariuki Njenga2,*

Departments of Veterinary Diagnostic Medicine1 and Veterinary PathoBiology,2 College of Veterinary Medicine, University of Minnesota, St. Paul, Minnesota 55108, and Southeast Poultry Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Athens, Georgia 306053

Received 14 April 2000/Returned for modification 28 July 2000/Accepted 27 August 2000

The matrix (M) protein of avian pneumovirus (APV) was evaluated for its antigenicity and reliability in an enzyme-linked immunosorbent assay (ELISA) for diagnosis of APV infection, a newly emergent disease of turkeys in United States. Sera from APV-infected turkeys consistently contained antibodies to a 30-kDa protein (M protein). An ELISA based on recombinant M protein generated in Escherichia coli was compared with the routine APV ELISA that utilizes inactivated virus as antigen. Of 34 experimentally infected turkeys, 33 (97.1%) were positive by M protein ELISA whereas only 18 (52.9%) were positive by routine APV ELISA 28 days after infection. None of the serum samples from 41 uninfected experimental turkeys were positive by M protein ELISA. Of 184 field sera from turkey flocks suspected of having APV infection, 133 (72.3%) were positive by M protein ELISA whereas only 99 (53.8%) were positive by routine APV ELISA. Twelve serum samples, which were negative by M protein ELISA but positive by routine APV ELISA, were not reactive with either recombinant M protein or denatured purified APV proteins by Western analysis. This indicates that the samples had given false-positive results by routine APV ELISA. The M protein ELISA was over six times more sensitive than virus isolation (11.5%) in detecting infections from samples obtained from birds showing clinical signs of APV infection. Taken together, these results show that ELISA based on recombinant M protein is a highly sensitive and specific test for detecting antibodies to APV.

* Corresponding author. Mailing address: Department of Veterinary PathoBiology, University of Minnesota, 1971 Commonwealth Ave., St. Paul, MN 55108. Phone: (612) 625-2719. Fax: (612) 625-5203. E-mail: Njeng001{at}tc.umn.edu.

Journal of Clinical Microbiology, November 2000, p. 4010-4014, Vol. 38, No. 11
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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Antimicrob. Agents Chemother. Clin. Microbiol. Rev.
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Copyright © 2000 by the American Society for Microbiology. All rights reserved.