Designing a competitive ELISA for evaluation of anti-snake venom serum potency

Document Type : Full Research Paper

Authors

1 Dept. of Biochem & Proteomics, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.

2 Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

3 Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.

4 Dept. of Serum & Proteomics, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.

5 Dept. of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.

Abstract

The venom of Naja naja oxiana, Vipera lebetina, Vipera albicornouta, Echis carinatus, Pseadocerastus persicus and Agkistrodon halys snakes are used to determine the potency of the polyvalent sera in neutralizing the venom. LD50 value per mouse was determined 6.5 μg. Neutralizations methods for obtaining potency of anti venom are competitive ELISA (Enzyme linked immunosorbent assay) and in vivo assay (ED50). Both of these tests were performed to estimate the serum potency in 15 samples of hyper-immune equines. Results of competitive ELISA showed that 1/12000 diluted serum can inhibits 50 percent of antigens. Competitive ELISA was compared with current biological assay ED50. Significant correlations between ELISA titers and values of ED500 at level of P≤ 0.01 and r=0.95 was observed, that indicates competitive ELISA can estimate antibody neutralizing capacity of the serum as well as the in vivo assay. The results of present study shows C-ELISA that measures Ag-Ab complex can used as a suitable replacement method for lethal neutralizing in vivo method.

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References

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History

  • Receive Date: 23 July 2015
  • Revise Date: 30 January 2016
  • Accept Date: 23 July 2015

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