STANDARDIZATION AND EVALUATION OF ANTIBODY RESPONSE BY PLATE-ELISA FOR THE DETECTION OF PARAMPHISTOMOSIS IN RUMINANTS

Disease transmission, diagnosis and preventive measures have become a major concern for the scientists working in national and international institution throughout the world. Paramphistomosis is a parasitic disease occurring in domestic ruminants causing economic loss to livestock industry. Early diagnosis of the disease is very important so that the loss due to disease can be curtailed by the appropriate treatment. Immunodiagnostic assays are helpful in the diagnosis of parasitic diseases. Hence immunological tests especially microtitre plate enzyme linked immunosorbant assay (ELISA) are the mainstay of diagnosis. Enzyme linked immunosorbant assay (ELISA), one of the immunodiagnostic assays is helpful in the diagnosis of paramphistomosis. More than 500 clinical/field sera samples of buffaloes, goats, sheep and cattle were collected from Bareilly, Delhi, Dehradun and Ludhiana under DST (GOI) sponsored project. Microtitre plate (Greiner) containing 96 well for Indirect Plate-ELISA was standardized for the detection of anti-Paramphistomunepiclitum antibodies in immunized rabbit using somatic antigen of P.epiclitum ranging from 1µg/ml to 10µg/ml, HRPO conjugate dilutions 1:1000 to 1:8000 and the range of sera dilution from 1:50 to 1:1,60,000. The optimum concentration of adult somatic P. epiclitum antigen was observed to be 2µg/ml, conjugate dilution 1:1000 and sera dilution at 1:200 and 1:400. A total of 222 clinical/field sera samples of ruminants were tested. Out of the total 106 ruminant samples were found to be positive with the incidence rate of 47.75 %. The highest percent positivity (84.0%) was found in naturally infected buffaloes followed by 26.25% in goats and 12.5% in sheep by indirect plate-ELISA. The observations on sensitivity and specificity of plate-ELISA test and immune response of antiP.epiclitum antibodies in experimental animal were also evaluated. The observation of high titre during the study is very effective for the detection of anti-P.epiclitum antibodies in field survey or in clinical cases. It is also helpful in the characterization of immunodominant antigens for the immunological control of the disease. Hence, indirect plateELISAis very important for the detection of paramphistomosis in domestic ruminants in early stages.