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<< JRScience Home     Volume 15 No.4    December, 2004    << Back to Volume Home

 

STANDARDIZATION OF COMPETITIVE ELISA FOR MEASUREMENT OF ANTIBODIES PRODUCED AGAINST Staphylococcus aureus

 

Khalil-ur-Rehman1, M. Anjum Zia2,*, Bushra Parveen1, Shagufta Kamal1 and M. Arshad2

1Department of Chemistry (Biochemistry) and 2Department of Veterinary Microbiology, University of Agriculture, Faisalabad, Pakistan.

Email: rmazia@hotmail.com

 

Abstract: Formalin, heat killed Staphylococcus aureus was prepared by growing on staph-110 agar medium. Different doses of antigen were injected in rabbits and after 14 days collected the serum. Antibodies isolated by ammonium sulfate precipitation, protein contents were estimated through Biuret as well as UV methods. The antibodies were conjugated with 10 mg soybean peroxidase having activity 83.02 U mL-1 using two-step glutaraldehyde method. Conjugation was detected by competitive ELISA and best results were recorded at 1:200 dilution of the conjugates.   

 

Keywords: Antibodies, ELISA, rabbits, Staphylococcus aureus.