Detection of anti–<i style="">Haemonchus contortus</i> antibodies in sheep by dot- ELISA with immunoaffinity purified fraction of ES antigen during prepatency

Abstract

Haemonchosis is a very common disease in small ruminants caused by <i style="">H. contortus,</i> a blood sucking parasite causing anaemia that may be fatal particularly to young animals. Therefore, detection of the infection during prepatent period is important for early treatment. Excretory-Secretory (ES) protein of <i style="">H. contortus</i> was purified through immunoaffinity chromatography. Dot -ELISA was performed with crude ES antigen as well as immunoaffinity purified fraction (F-1) with experimental and natural sera of sheep infected with <i style="">H. contortus</i>. Solid dot formation took place with 4 day, 1, 2 and 3 weeks post infection sera. Dot formation did not take place with negative control serum and uninfected control animal serum. When crude ES antigens was reacted to natural sheep sera having <i style="">H. contortus</i> infection, 60% sera samples showed solid dot formation whereas in F-1 fraction 75% of the sera samples showed solid dot indicating purified fraction was a more potent antigen. Crude ES and F-1 were also fractionated through SDS-PAGE. ES antigen revealed polypeptides in the range of 10 to 200 kDa of which 26, 32, 60 and 120 kDa were found more prominent. F-1 fraction on SDS-PAGE analysis revealed only four polypeptides of 26, 32, 60, and 120 of which 60 and 120 kDa were found to be most prominent. Results indicate that the purified fraction of ES antigen may be utilized for early diagnosis of haemonchosis. Further studies on cross antigenicity of this fraction with other nematode and trematode needs to be conducted.