Detection of Paramphistomum cevi Antibodies in Rabbits using ELISA and DID
Tariq Ahmed1, M. Z. Chisti1, Syed Tanveer1, Abdul Baqui1 and Zaffar Amin Shah2*1P.G. Department of Zoology Kashmir university. 2Sheri Kashmir Institute of Medical Science (Department of Immunology) (India).
Abstract: Sheep (Ovis aries) is infected with a variety of gastrointestinal helminths , of which notably Fasciola hepatica and Paramphistomum cervi are prominent and pathogen causing a lot of morbidity and mortality (Mukherjee and Chauhan, 1965). Immature stages of P. cervi are highly pathogenic (Horak 1971) but the routine parasitological diagnosis is difficult, under the circumstances immunological test applied to the present study, is aimed at investigating the relative immunodiagnostic reliability and sensitivity of the Ouchterlony gel diffusion test and ELISA in paramphistomiasis. Sheep blood was collected at the local abattoir naturally infected with Paramphistomum cervi. Using somatic antigen of whole worms derived from P.cervi was prepared by homogenisation, sonication and centrifugation at 10,000 r pm for 20 minutes at 4oC. Rabbits were immunized with the antigen mixed with Freunds complete adjuvant (1:1 ratio) for raising hyperimmune sera. Blood was collected at regular intervals by puncturing ear vein of the rabbit. The naturally infected sheep sera were also collected from the slaughter houses. The Ouchterlony test and ELISA were found to be positive as early as 2-4th weeks post-infection. By gel diffusion test, two precipitation bands were observed, and at 6th – 8th weeks of post infection by ELISA. An indirect ELISA standardized for detection of anti-Paramphistomum antibodies using antigen concentration of 2µg / ml was used on coating buffer. Indirect ELISA revealed antibody titre as high as 1: 12,800 in rabbit sera where as in sheep sera 1: 6400.
Keywords: P. cervi; Indirect ELISA; Freunds Complete Adjuvant (FCA); Immunodiagnosis; Sheep; Rabbit Back to TOC