Designing an ELISA system to measure the antibody produced in guinea pigs immunized with FMD virus

Document Type : Full Research Paper

Authors

1 Department of Biology, North Tehran Branch, Islamic Azad University, Tehran, Iran

2 Department of Microbiology, School of Specialized Science, Science and Research Branch, Islamic Azad University, Tehran, Iran

3 Department of Biotechnology, Razi Vaccine and Serum Research institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran

Abstract

Foot-and-mouth (FMD) disease is an infectious and fatal viral disease that affects cloven-hoofed animals. Although different biological and serological approaches are still applied to study this disease, ELISA test based on the distinct format, antigen type and specific antibody reinforce its predominance in different research areas of FMD, and this may replace the traditional methods in the near future. This study was conducted to design an ELISA system to measure the antibody produced in guinea pigs immunized with FMD virus. The purified FMD virus was obtained and then protein concentration of FMD solution was measured. Immunization of 5 guinea pigs was done and then blood sample was taken for obtaining serum. Ouchterlony test was used and finally the ELISA system was designed. Protein concentration was 3.5 mg/ml. By conducting ouchterlony test, the sediment lines between the serum and the antigen of the FMD virus were formed. Using ELISA and optimizing the conditions, the optimal concentration of antigen was 1 μg/well and a serum dilution of 1: 100 was achieved. In the ELISA system, the standard deviation was 0.35 and the cutoff value was 1.03, So that, the samples above absorbance of 1.03 are positive and the samples less than absorbance of 1.03 are negative. The in house ELISA can be used to measure the antibody produced in guinea pigs immunized with FMD virus, which can using for suitable and cost effective laboratory diagnostic tests.

Keywords

References

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Veterinary Research & Biological Products
Volume 34, Issue 3
October 2021
Pages 13-21

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History

  • Receive Date: 30 December 2019
  • Revise Date: 19 April 2020
  • Accept Date: 21 April 2020

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