SIGIT SETYONO RAHARJO, 060333114 (2009) DIAGNOSA BRUCELLOSIS DENGAN MILK RING TEST DAN ELISA DI KOPERASI UNIT DESA "DADI JAYA" PURWODADI KABUPATEN PASURUAN. Skripsi thesis, UNIVERSITAS AIRLANGGA.

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Abstract

The purposes of this research is to diagnoses Brucellosis through milk samples with MRT and indirect ELISA method, and then compare both the result test of them. As the research method, milk samples has taken as much as 10 ml from each milk can of farmer who comes to KUD "Dadi Jaya", then it's carried to the laboratory for MRT and ELISA procedure. Before doing the ELISA procedure, milk sample must be centrifuge at a velocity that enables the separation between the lipid layer and lactoserum, called milk plasma. It consists of proteins and lactose, without lipid, because lipid could make ELISA procedure has failed. The principal of Indirect ELISA is LPS of Brucella sp. which coated in a well of microplate will bind with antibodies that come from milk plasma. Anti-bovine Ig G peroxides conjugate have a function as the secondary antibody, and then forming a complex reaction: (Ag)—(Ab)–(anti-bovine Ig G peroxides). The buffer peroxides substrate is important to product a coloured after that reaction. The Result of ELISA could be read using ELISA reader at ?, = 450 run. The Optical Density values are important to classified positive or negative Brucellosis, through the value of the positive control. The principal of MRT is milk sample towards MRT antigen. Antibodies present in the milk will bind to the antigen from MRT reagent, and then agglutination of the lipid-grains happened. The agglutinin makes the lipid-grains float, and then become blue ring form at the milk surface as a positive Brucellosis sample. It's because the blue colour which's produced MRT antigen. The results has shown that 5 of 60 milk samples are positive Brucellosis according to Milk Ring Test, but only one sample are positive by using the ELISA methode.

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