Molecular identification of Escherichia coli K99 and evaluation of the serum antibody titer in laboratory animals
Subject Areas : Immunology
Faezeh Farhani
1
,
Yahya Tahamtan
2
,
Mohammad Kargar
3
1 - 1M.Sc., Department of Microbiology, Jahrom branch, Islamic Azad University, Jahrom, Iran.
2 - Associate Professor, Department of Microbiology, Razi Vaccine and Serum Research Institute, Shiraz, Iran
3 - Associate Professor, Department of Microbiology, Jahrom branch, Islamic Azad University, Jahrom, Iran.
Keywords: ELISA, Escherichia coli, Calves, Fimbriae K99, Diarrhea,
Abstract :
Background & Objectives: Escherichia coli is one of the primary etiologic agents for diarrhoea in neonatal calves, and the maternal immunization is an effective protection for the neonates because of feeding with colostrum. This study was aimed to molecular detection of E. coli K99 and evaluation of the serum antibody titers in laboratory animals. Materials & Methods: This study was experimentally carried out on rat to detect the marker genes. First, a Multiplex PCR was applied to detect the Sta, F41 and K99 pathogenic genes. Following growth of E. coli K99 bacteria into Minca broth, the bacteria were inactivated by 0.4% formaldehyde. The antigen was prepared after washing with PBS and centrifugation. The rats were subcutaneously inoculated with inactivated antigen with 107 and 109 CFU/ml in two groups. The control group was inoculated with a suspension containing all ingredients except for the antigen. Rat was allowed to nurse immediately after injection, and blood samples were taken for serum titration for 8 weeks. Results: An increase in the titer of serum antibody was seen after four weeks, and it continued after seven weeks. The antibody titration of rate was significantly higher in the immunized rat group than in the control group. We found good correlation in two groups. There is a grateful increase in antibody titer in group 2 (received 109 CFU/ml antigen) than in group 1 (received 107 CFU/ml antigen), but decreasing rate in the both groups were the same. Conclusion: These findings indicate that the use of killed antigen with high titer preparation containing sufficient antigen and may provide passive protection in animals. This resulted in by stimulation of immune cell due to high dose antigen.