عنوان مقاله [English]
The major agents cause abortion in cattle are Neospora caninum, Infectious Bovine Rhinotracheitis (IBR), and Bovine Viral Diarrhea (BVD). These infection agents cause reproductive disorders which can lead in economic loss in dairy and beef herds. Serological test including ELISA are calculated as the best diagnostic ways for Neospora caninum, IBR, and BVD. The aim of this study was to develop a multiplex Dot-ELISA kit for simultaneous detecting of three abortion agents: Neospora caninum, IBR, and BVD. The blood samples were collected from 184 cows with a history of abortion in the northern of Fars province. Sera were analyzed by Multiplex Dot-ELISA and commercial ELISA kits, and the results were compared to each other. The commercial kit showed 32%, 98,9% and 64.1% positive results for Neospora caninum, IBR, and BVD, respectively. On the other hand, positive results for Multiplex Dot-ELISA showed 29.3%, 93.5% and 79.3% for these agents. Kappa agreement assessment demonstrated that appropriate agreement between multiplex Dot-ELISA results and ELISA kits, and McNemar test showed that both methods have the same efficacy for the identification of these abortion agents. The results of this study revealed that multiple Dot-ELISA kit, could be a simple and economical method for the simultaneous detection of three abortion agents in cattle. This experimentally kit is designed as the first step in designing a multiple kit for the diagnosis of infections in cows which could be introduced with complementary studies as a commercial kit in the future.
1. Alenius, S., Lindberg, A., Larsson, B. 1997. A national approach to the control of bovine viral diarrhoea virus. Proc. of the third ESVV symposium on the control of pestivirus infections, Lelystad. The Netherlands, 162-169.
2. Bashir, S., Rashmi, Singh, R., Sharma, B., Sharad, K., Yadav, AK. 2011. Development of a sandwich ELISA for the detection of bovine herpesvirus type 1. Asian Pacific Journal of Tropical Medicine, 4: 363-366.
3. Baszler, T., Adams, S., Vander-Schalie, J., Mathison, B., Kostovic, M. 2001. Validation of a commercially available monoclonal antibody-based competitive-inhibition enzyme-linked immunosorbent assay for detection of serum antibodies to Neospora caninum in cattle. Journal of Clinical Microbiology, 39: 3851-3857.
4. Bjorkman, C., Uggla, A. 1999. Serological diagnosis of Neospora caninum infection. International Journal of Parasitology, 29: 1497-1507.
5. Blanco, Rafael D., Cintia, F., Fidelis, Leandro, S., Araujo, Adriana, M., Henao, Jose, A., Cardona, and etal. 2014. Development and standardization of Dot-ELISA using recombinant peptides for serological diagnosis of Neospora caninum. Pesquisa Veterinary Brasilia, 34: 8.
6. Dubey, J.P., and Schares, G. 2006. Diagnosis of Bovine Neosporosis. Veterinary Parasitology, 140: 1-34.
7. Graham, D.A., Mawhinney, K.A., McShane, J., Conno,r J.A., Adair, B.M., Merza, M. 1997. Standardization of enzyme-linked immunosorbent assays (ELISAs) for quantitative estimation of antibodies specific for infectious bovine rhinotracheitis virus, respiratory syncytial virus, parainfluenza-3 virus, and bovine viral diarrhea virus. Journal of Veterinary Diagnostic Investigation. 9: 24-31.
8. Crowther, J.R. 2000. The ELISA Guidebook: Methods on Molecular biology. Second edition. Human press, Totowa, Jersey, 85-140.
9. Hamidinejat, H., Haji hajikolaei, M., Ghorbanpoor, M., Namavari, M., and Mohamad Ali Gol, S. 2013. Development and Standardization of Dot – ELISA for Detection of Neospora caninum Antibodies in Cattle and Comparison with Standard Indirect ELISA and Direct Agglutination Test (DAT). Iran Journal of Parasitology, 8:634–640.
10. Hemat Zadeh, Farhid. 2001. Development of an immunofluorescent kit for IBR serodiagnosis. Iranian Journal of Veterinary Medicine 56, no. 2 (In persian).
11. Hemat Zadeh, Farhid. 2003. Development of a dot Elisa kit for IBR serodiagnosis in cattle. Iranian Journal of Veterinary Medicine, 56, no. 6 (In Persian)
12. Hemmatzadeh, F., Amini, F. 2009. Dot-blot enzyme immunoassay for the detection of bovine viral diarrhea virus antibodies. Veterinarski Arhiv, 79: 343–350
13. Hoar, B.R., McQuarry, A.C., Hietala, S.K. 2007. Prevalence of Neospora caninum and persistent infection with bovine viral diarrhea virus in dairy-breed steers in a feedlot. Journal of American Veterinary Medical Association, 230: 1038–1043.
14. Hudson, l., Hay, FC. 1989. Practical immunology, 3th edition. Blackwell scientific publication, 15-42
15. Itoh M1, Sato S.1990. Multi-dot enzyme-linked immunosorbent assay for serodiagnosis of trematodiasis. Southeast Asian J Trop Med Public Health. 21:471-4.
16. Jones, C., Newby, TJ., Holt, T., Doster, A., Stone, M., Ciacci-Zanella, J., Webster, JC., Jackwood, MW. 2000. Analysis of latency in cattle after inoculation with a temperature sensitive mutant of bovine herpesvirus 1 (RLB106), Vaccine, 18: 3185–3195.