Preparation of Antiserum against Salmonella O Antigen for application in Agglutination and Elisa tests

Introduction and Objectives: Salmonella infection (salmonellosis) is a common bacterial disease that affects the intestinal tract and usually caused by eating raw or undercooked meat, poultry, eggs or egg products. A few varieties of these bacteria result in typhoid fever, which is more common in developing countries. Several methods like Multiplex or real-time PCR, Elisa, and Agglutination are used to identify these bacteria. However, normally rapid, cost effective and easy diagnostic methods such as agglutination test is recommended. In Iran, positive control antiserum used in diagnostic kits works based on polyvalent agglutination and are against O and H antigens. The purpose of this research was to produce specific anti-sera against O antigen for using in agglutination and ELISA kits. Materials and Methods: New Zealand white rabbits were immunized by intravenous injections of inactivated bacterial O antigen adjusted to a cell density equivalent to a turbidity of a McFarland number 3 standard. Serum collection was performed 7 days after last injection. Collected Antisera was tested with positive human specimens as well as cross-reaction antibodies. Absorption method was used to obtain specific anti-sera against O antigen. Produced Anti-O antibody was mixed with bacterial H antigen and incubated for 1 hours in 37˚c. The Mixture was centrifuged and supernatant was collected. Results: Results showed high quality and quantity of mono-specific antibody, which showed no cross-reaction with H antigen. Conclusion: Agglutination based assays are rapid, cost effective and easy for primary screening of salmonella infections. In addition, anti-sera can be used in Elisa and Immunofluorescence kit after purification.

کلیدواژه ها:

Salmonella ، O antigen ، specific antiserum ، agglutination ، ELISA