Development of ELISA kit for the diagnosis of sheep brucellosis
2004
Ghasemipour, Navid | Mostafae, Ali | behrozikhah, Mohammad Ali
Brucellosis is one of the most important zoonotic diseases that is caused by several Brucella species. This disease is noticeable from the viewpoint of economic (abortion, decreased milk and fertility) and sanitary (transfer to human).An accurate diagnosis of brucellosis based on clinical signs is not always possible. Isolation of the causative agent is the most accurate test for the detection of brucellosis. Besides false negative, the other disadvantage of bacterial isolation has been reported to be non costeffective. In the absence of bacterial isolation, other assays such as different serological tests are used widely. Among serological assays, ELISA has been known as one of the convenient methods to detect the disease. The aim of this research was to develop an ELISA kit for the detection of brucellosis in sheep, using lipopolysaccharide (LPS) and whole cell of killed Brucella melitensis variety of ١٦M. LPS extracted as described by Morrison and Leive. Sheep IgG was purified, using ion exchange chromatography and then injected to ٤ rabbits for raising antisheep IgG in rabbits. The prepared anti serum after purifying was conjugated with peroxidase and next, ELISA assay was set up. The cutoff value was calculated as the mean specific OD of control sera plus ٣ standard deviations. The sensitivity and specificity of developed kit for LPS and whole cell were ١٠٠ and ٨٠ percent and ١٠٠ and ١٠٠ percent, respectively.
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