Use of Nickel Chloride in Dot-ELISA for Detection of Haemonchus sp. Infection in Sheep

Abstract

Haemonchosis is a major cause of production losses in livestock, particularly sheep and goat. In this study, Nickel chloride is used in dot-ELISA for the detection of infection in sheep, experimentally-infected with 10,000 L 3 Haemonchus contortus. Serum samples from infected sheep were collected at weekly intervals. Adult somatic antigen was prepared from the worms collected from the local abattoir at Meerut (U.P.). Nitrocellulose membranes (0.2 ?m) were dotted with 10 ?l antigen that contained 8-10 ?g protein, and were incubated at 37°C for 2 h. Non-specific binding sites were blocked with blocking agent casein in PBS( phosphate-buffered saline). Positive, negative and uninfected control sera from sheep, both experimentally- and naturally-infected, were used. Positive reaction with blue-black solid dot formation was obtained with the sera, as early as first week post-infection, with immunoaffinity- purified adult somatic antigen.

Keywords: dot-ELISA, Haemonchus contortus, immunoaffinity chromatography, Nickel chloride.