Document Type : Original Article


1 Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Hamedan, Iran

3 Department of Pathobiology, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran

4 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

5 Quality Control Department, Razi Vaccine and Serum Research Institute, Agricultural Education and Extension Research Organization (AREEO), Karaj, Iran

6 Department of Food Hygiene, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

7 Department of Pharmacognosy, Faculty of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran


Diagnosis of bovine viral diarrhea (BVD) relies on the detection of antibodies against its viral causing agent, bovine viral diarrhea virus (BVDV). Here, we designed a novel competitive ELISA (cELISA) using the most immunogenic part of BVDV nonstructural protein 3 (NS3), as a single ELISA recombinant antigen, along with a monoclonal antibody to detect antibodies against BVDV in sera of infected animals. Hence, 197 serum samples were tested by this cELISA and the results were compared to the results obtained from virus neutralization test (VNT) as the gold standard method for diagnosis of BVD. McNemar’s test indicated that there was no significant difference between the results of this newly designed cELISA and VNT. Meanwhile, kappa coefficients showed that there was a high correlation between these two assays. The relative sensitivity and specificity of cELISA with respect to VNT were 93.90% and 100%, respectively, suggesting that this newly designed cELISA could be a useful diagnostic tool for detection of BVDV infection. Moreover, as NS3 is highly conserved among Pestiviruses and the developed ELISA is a competitive one, it could potentially be applied to detect BVDV infection in other domestic and wildlife species.


  1. Baker JC. The clinical manifestations of bovine viral diarrhea infection. Vet Clin North Am: Food Anim Pract 1995; 11(3): 425-445.
  2. Greiser-Wilke I, Grummer B, Moennig V. Bovine viral diarrhea eradication and control programmes in Europe. Biologicals 2003; 31(2): 113-118.
  3. Presi P, Heim D. BVD eradication in Switzerland--a new approach. Vet Microbiol 2010; 142(1-2): 137-142.
  4. Collett MS, Wiskerchen M, Welniak E, et al. Bovine viral diarrhea virus genomic organization. Arch Virol Suppl 1991; 3: 19-27.
  5. Gorbalenya AE, Koonin EV, Donchenko AP, et al. Two related superfamilies of putative helicases involved in replication, recombination, repair and expression of DNA and RNA genomes. Nucleic Acids Res 1989; 17(12): 4713-4730.
  6. Wiskerchen M, Collett MS. Pestivirus gene expression: protein p80 of bovine viral diarrhea virus is a proteinase involved in polyprotein processing. Virology 1991; 184(1): 341-350.
  7. Chimeno Zoth S, Taboga O. Multiple recombinant ELISA for the detection of bovine viral diarrhoea virus antibodies in cattle sera. J Virol Methods 2006; 138(1-2): 99-108.
  8. Bolin SR, Ridpath JF. Specificity of neutralizing and precipitating antibodies induced in healthy calves by monovalent modified-live bovine viral diarrhea virus vaccines. Am J Vet Res 1989; 50(6): 817-821.
  9. Donis RO, Corapi WV, Dubovi EJ. Bovine viral diarrhea virus proteins and their antigenic analyses. Arch Virol Suppl 1991; 3: 29-40.
  10. Sandvik Laboratory diagnostic investigations for bovine viral diarrhoea virus infections in cattle. Vet Microbiol 1999; 64(2-3): 123-134.
  11. Lecomte C, Pin JJ, De Moerlooze L, et al. ELISA detection of bovine viral diarrhoea virus specific antibodies using recombinant antigen and monoclonal antibodies. Vet Microbiol 1990; 23(1-4): 193-201.
  12. Vanderheijden N, De Moerlooze L, Vandenbergh D, et al. Expression of the bovine viral diarrhea virus Osloss p80 protein: its use as ELISA antigen for cattle serum antibody J Gen Virol 1993; 74(Pt 7): 1427-1431.
  13. Reddy JR, Kwang J, Okwumabua O, et al. Application of recombinant bovine viral diarrhea virus proteins in the diagnosis of bovine viral diarrhea infection in cattle. Vet Microbiol 1997; 57(2-3): 119-133.
  14. Mahmoodi P, Shapouri MR, Ghorbanpour M, et al. Epitope mapping of bovine viral diarrhea virus nonstructural protein 3. Vet Immunol Immunopathol 2014; 161(3-4): 232-239.
  15. World Organization for Animal Health (OIE), Manual of diagnostic tests and vaccines for terrestrial animals 2021, Chapter 3.4.7, Available at: Accessed July 11, 2022.
  16. Edwards S. The diagnosis of bovine virus diarrhea-mucosal disease in cattle. Rev Sci Tech 1990; 9(1): 115-130.
  17. Deregt D, Dubovi EJ, Jolley ME, et al. Mapping of two antigenic domains on the NS3 protein of the pestivirus bovine viral diarrhea virus. Vet Microbiol 2005; 108(1-2): 13-22.
  18. Bhatia S, Sood R, Mishra N, et al. Development and evaluation of a MAb based competitive-ELISA using helicase domain of NS3 protein for sero-diagnosis of bovine viral diarrhea in cattle and buffaloes. Res Vet Sci 2008; 85(1): 39-45.