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 Gulati, B. R.
Right arrow Articles by Njenga, M. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gulati, B. R.
Right arrow Articles by Njenga, M. K.

 Previous Article  |  Next Article 

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.


This article has been cited by other articles:

  • Hamelin, M.-E., Boivin, G. (2005). Development and Validation of an Enzyme-Linked Immunosorbent Assay for Human Metapneumovirus Serology Based on a Recombinant Viral Protein. CVI 12: 249-253 [Abstract] [Full Text]  
  • Luo, L., Sabara, M. I., Li, Y. (2005). Expression of Recombinant Small Hydrophobic Protein for Serospecific Detection of Avian Pneumovirus Subgroup C. CVI 12: 187-191 [Abstract] [Full Text]  
  • Alvarez, R., Njenga, M. K., Scott, M., Seal, B. S. (2004). Development of a Nucleoprotein-Based Enzyme-Linked Immunosorbent Assay Using a Synthetic Peptide Antigen for Detection of Avian Metapneumovirus Antibodies in Turkey Sera. CVI 11: 245-249 [Abstract] [Full Text]  
  • Turpin, E. A., Lauer, D. C., Swayne, D. E. (2003). Development and Evaluation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Avian Metapneumovirus Type C-Specific Antibodies in Multiple Domestic Avian Species. J. Clin. Microbiol. 41: 3579-3583 [Abstract] [Full Text]  
  • Shin, H.-J., Cameron, K. T., Jacobs, J. A., Turpin, E. A., Halvorson, D. A., Goyal, S. M., Nagaraja, K. V., Kumar, M. C., Lauer, D. C., Seal, B. S., Njenga, M. K. (2002). Molecular Epidemiology of Subgroup C Avian Pneumoviruses Isolated in the United States and Comparison with Subgroup A and B Viruses. J. Clin. Microbiol. 40: 1687-1693 [Abstract] [Full Text]  
  • Gulati, B. R., Munir, S., Patnayak, D. P., Goyal, S. M., Kapur, V. (2001). Detection of Antibodies to U.S. Isolates of Avian Pneumovirus by a Recombinant Nucleocapsid Protein-Based Sandwich Enzyme-Linked Immunosorbent Assay. J. Clin. Microbiol. 39: 2967-2970 [Abstract] [Full Text]  
  • Shin, H.-J., Njenga, M. K., McComb, B., Halvorson, D. A., Nagaraja, K. V. (2000). Avian Pneumovirus (APV) RNA from Wild and Sentinel Birds in the United States Has Genetic Homology with RNA from APV Isolates from Domestic Turkeys. J. Clin. Microbiol. 38: 4282-4284 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»