Standardization of Dot-ELISA for diagnosis of brucellosis in sheep and comparison of it with other conventional serological tests

Authors
Maryam Abbasvali
Seyed Mohammad Hossein Hosseini
Roya Firouzi
Abstract
In this study, using Brucella antigen, prepared by Vaccine and Serum Research Institute (karaj, Iran), anti-sheep IgG antibody conjugated with peroxidase and 4- chloronaphtol as a coloring agent and hydrogen peroxideas substrate, the Dot-ELISA test for the diagnosis of ovine brucellosis was standardized in such a way that with appropriate dilution of serum, pooled negative sera gave negative and pooled positive sera gave positive reaction. Then, on 100 positive and 10 negative sheep serum samples various conventional serological tests such as Rose Bengal Plate Test (RBPT), Standard Tube Agglutination Test (STAT), 2-Mercaptoethanol test (2ME) and standardized Dot-ELISA test, were applied. Out of positive sheep serum samples, 100, 91, 93 and 100 percentage were found to be positive with RBPT, STAT, 2ME, and standardized Dot-ELISA test, respectively. All the negative samples were detected negative in all four tests. The resultsshowed that standardized Dot-ELISA test has 100%, 92% and 94% agreement rate with RBPT, STAT and 2ME tests, respectively (p< 0.001) and with more sensitivity is also rapid, simple and reliable to be used as a field test for the serodiagnosis of suspected ovine samples.
Keywords
Brucellosis-Dot-ELISA-Sheep-Standardization-