Evaluation of Different Antigens in Western Blotting Technique for the Diagnosis of Sheep Haemonchosis

Abstract

Background: To use different methods for serodiagnosis of ruminants' haemonchosis is important because detection of egg in the faeces is not so reliable.

Methods: Peptide bands of 5 different crude antigens of intestine, uterus, cuticle, whole male and whole female of Hae­monchus contortus were determined using SDS-PAGE and western blotting. Five lambs were infected with 10,000 third stage larvae of H.contortus and 2 parasites free were kept as control.  Positive and negative sera collected from infected and no infected animal were tested using western blotting for immunodiagnostic antigens.

Results: In electrophoresis the major peptide bands of crud antigens of uterus, intestine, cuticle, whole male and whole fe­male of H. contortus were 7, 2, 6, 8 and 5, respectively at molecular weights of 15 to 110 kDa. In immunoblotting positive and negative sera were compared and the molecular weight of specific protein bands for Haemonchus in sheep was deter­mined. Two major peptide bands belong to intestine and uterus with 35 and 40 kDa molecular weight, respectively, were specific for diagnosis of the parasite infection.

Conclusion: Our findings suggest the use of these antigens with different molecular weights for immunodiagnosis of hae­monchosis in sheep as a primary screening test is promising.