Development and preparation of anti-Human globulin (IgG) for detection of brucellosis in Coombs Wright test

Introduction and Objectives: Brucellosis is a common illness in man and livestock which is caused by the Brucella Species. Animal health is crucial for us for dairy consumption. An important diagnostic test for this bacterium is the Wright test. Because IgG is not a strong agglutinin with its monomeric structure, false negative results of Wright test is probable and therefore, it is better to use the Coombs wright test to detect this type of antibody. Due to the importance of the presence of IgG antibodies in the chronic stages of the disease, and the negativity of the wright tube test, we use anti-human globulin. The purpose of this study was to prepare antiserum against IgG immunoglobulin for use in the coombs Wright test. Materials and Methods: 250 μg of purified human IgG was mixed with one milligram of alum adjuvant. New Zealand White Rabbit was injected for four times with a week s intervals. Blood collection and serum separation were performed and purification was done using ammonium sulfate precipitation, Tangential flow filtration and DEAE ion exchange chromatography methods. Results: The results after the completion of anti-human globulin purification steps showed that by using spectrophotometric method and the Lowry and Bradford protein assay methods, as well as the SDS-Page Gel, the purity of the isolated anti-human globulin was quantitatively and qualitatively acceptable. Conclusion: Purified antibodies against IgG can be immunogenic in the clinical diagnosis of Coombs-Wright tests, the cross-match as well as the detection of blood groups with weak Rh or Du.

کلیدواژه ها:

Brucellosis ، Immunoglobulin IgG ، Coombs Wright