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Development And Validation of An In-House Brucella Smooth Lipopolysaccharide as A Coating Antigen in ELISA For Serodiagnosis of Bovine Brucellosis

of Smooth Lipopolysaccharide as A Coating Antigen in ELISA For Serodiagnosis Abstract Background: Brucellosis is an important zoonotic bacterial disease of global health importance affecting different animals and man. Brucellosis control and eradication procedures are highly depending on accurate diagnostic tools and effective and safe vaccination programs. Indirect ELISA, complement fixation tests, Buffered Acidified Plate Agglutination and the Rose Bengal plate agglutination are usually used for testing animals against bovine brucellosis. The aim of the work is to evaluate and validate the diagnostic efficacy of in-house prepared S-LPS as an IELISA coating antigen in comparison to commercial ELISA kit used for diagnosis of bovine brucellosis. Materials and methods: an Indirect Enzyme Linked Immunosorbent Assay (I-ELISA) was developed using in house produced and titrated Smooth Lipopolysaccharide (S-LPS) as coating antigen and compared with the commercial kit using 115 negative and positive bovine sera. The performance parameters for each test were estimated using Complement Fixation Test (CFT) as a gold standard test. Results: the diagnostic performance characteristics for both ELISA tests were estimated to evaluate the diagnostic efficacy of in-house S-LPS. Brucella antibodies were detected in 88 (76.52%), 90 (78.26%) and 76 (66.09%), by in house i-ELISA, commercial i-ELISA and CFT, respectively. agreement for the positive result between the developed and commercial I-ELISA was in 14 positive sample and in 23 negative samples. Relative sensitivity was (97.37%) for both ELISA assay, while the specificity was 64.1% and 58.97% for developed I-ELISA and commercial ELISA kit respectively. Conclusion: Results revealed that in house prepared, titrated, and validated S-LPS can be used as a coating antigen for IELISA test for bovine brucellosis diagnosis as a sensitive test. On other hand- in house test is cheaper than commercially available kits and so, it could be used for wide surveillance program