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Development and Validation of the Indirect Whole Cell ELISA for Diagnosis of Bovine Brucellosis

A. Khogali, AE. O. Mohamed, M. Hussien, H. Elrufai

This study was carried out at autumn to diagnose Bovine Brucellosis using intact Brucella antigen (whole cell). Fifty blood samples were collected from Friesian cattle at Kuku Scheme (Khartoum North) from females above the age of puberty and haven’t been vaccinated against brucellosis before. The samples were tested with Rose Bengal to detect negative and positive sera then indirect ELISA tests using intact Brucella bacteria were carried out. Four known positive sera were used to choose optimum concentration of intact Brucella which will be used as an antigen. There was a great deal of variation amongst the tested sera in antibodies titer in which 30 samples were positive (40%) and 20 were negative with Rose Bengal and 4 were negative and 46 were positive (60%) with indirect ELISA. The result showed that the upper limit of the control sera is taken as the lower limit of positivity which called cut off point and took the mean and standard deviation and considered that negative sera were under 0.2 OD and the above of this were positive. In this study, Chi square test in SPSS program was used and showed the variation in detection of positive and negative samples between Rose Bengal and ELISA which used with intact bacteria with probability of >0.05 and this showed the significance of ELISA. The intact Brucella ELISA test showed high sensitivity and economic impact and detect all epitopes. This can be useful for diagnosis and epidemiological surveys, and may reduce the dependence on imported, expensive commercial materials.