Detection of antibodies against Trypanosoma evansi in dromnedary camels by ELISA using solubilized antigens

Abstract

An ELISA for detection of antibodies against Trypanosoma evansi in dromedary camels using 2 types of solubilized antigens. viz, Crude solubilized antigen (CSA) and detergebt solubilized antigen (DSA) was standardized using positive and negative control sera. The optimum conditions standardized for the test were coating the wells of micro-ELISA plate with 100 µl of solubilized antigens (at a concentration of 2 µg /ml), blocking with 5% skimmed milk powder in PBS-T (200 µl/well). rabbit anti-camel lg- HRPO conjugate diluted 1:300 to 1:1000 with PBS-T (100 µl/well) and colour development with freshly prepared substrate solution of OPD (200 µl/well). After measuring the absorbance value of positive and negative controls, test serum samples with twice the mean absorbance value of negative control sera were taken as positive and those below this value as negative. A total of 180 field collected serum samples of camels were screened with ELISA using CSA and DSA as test antigens and out of them 112 (62.22%) and 124 (68.88%) wer positive for T. easnsi antibodies respectively.