Study On The Rapid Detection Methods Of Animal Brucella

Brucellosis is one kind of amphixenosis, which is severely impaired in the world. The detection methods in my Country are bacteriology and serology (RBT and SAT). But. these methods have many drawbacks. It is necessary to establish new methods to improve detection level. This study established three methods to lay a foundation for Brucellosis control. To meet the demand of the quarantine purification of brucellosis, the papers study on the rapid detection method of brucellosis, the serological typing identification of pathogen detection antibody screening�'positive samples�'pathogens.1. Established colloidal gold immunochromatography assay(GICA) for the field detection brucella.According to double antigen sandwich detection antibody.we used SPA and bp26to set up the colloidal gold immunity chromatography test paper.The specific test showed the detection result of control serum were Correct.The sensitivity of test reached the level of SAT.The shelf life was6months in4℃.2. Established fluorescence quantitative PCR(FQ-PCR)for the quantitative detection brucella. According to the consensus sequence of brucella genome OMP10, we devised specific primer and Taqman fluorescent probe.We had cloned the gene part OMP10to carrier PMD19-T,extracted plasmid and prepared the standard substance and the standard curve.The result of specific testing showed that there was not fluorescence signal in compare bacterias.By the analysis of sensitivity,we heaved known that the lowest detection limit was10-4ng/μL.The threshold change was little in different test time and different reaction tubes in stability experiment and the coefficient of variability was less than2%.3.Established multiplex PCR for detection and classification of Brucella.According to multicopy insertion element IS711and its polymorphic site on lower reaches,we had devised primers of detection brucella and classification cow, sheep and pig brucella. The multiple PCR reaction system and condition had been optimized.The multiple PCR which was to detect and classificate the brucella of cow、sheep and pig had been set up. By detecting the comparing bacterial strain.there were not specificity amplification.This method was specificity.By analyzing the sensitivity.the method of multiple PCR could test the10-2ng/μL.