Preparation of diagnostic ELISA kits for detection of camelpox virus

Abstract

Peroxidase labeled immunoglobulins to camelpox virus (CPV) were prepared for use in various techniques of ELISA. Ten rabbits and three goats were inoculated with a mixture of camelpox virusand Freund’s adjuvant. Sera were pooled separately on the 10th day post the last inoculation and immunoglobulins were precipitated using saturated ammonium sulphate. The globulins were 2.8 g/dl and 2.5 g/dl for rabbits and goats respectively and used for peroxidase conjugation. The peroxidase labeled immunoglobulins were titrated and evaluated using direct solid phase ELISA, double antibody sandwich ELISA and dot immunoblot ELISA. The prepared conjugates gave specific and clear positive reactions till the dilution of 2000 and 1500 for rabbits and goats immunoglobulins respectively. The prepared labeled immunoglobulins could be successfully used in detection of camel pox viral antigen of local virulent and standard vaccinal strain of the virus using various ELISA techniques.

Keywords

• elisa; detection; camelpox virus