The Research On Omp19Antigenicity Of Brucella And Cross-Sectional Survey Of Human Brucellosis In Inner Mongolia

Brucellosis is a zoonotic infectious disease,the main source of infection is likestock such as sheep cow and so on,the sheep are more seriously.The effective means to control the disease are diagnosis and prevention.but so far,there is no way can be used to diagnose Brucellosis independently,but indirect enzyme-linked immunosorbent assay(iELISA) can be widely used, because of its simple operation and low experimental conditions requirement;Currently Brucellosis epidemic widely,Brucella have been isolated all over the country,when we want to identify the strain,we need the highly pathogenic standard strain as control,but it is diffic μLt to purchase and transport.there is also a big risk for staff;The agricultural and pastoral areas are the main concentrated outbreak regions,we can know the debelopmental trend of Brucellosis from on site investigation.So this research faces a study of Brucellosis from system diagnostics,biosaf ety and field investigations parties.The study one in order to improve the diagnosis of brucellosis,established the recombinated plasmid pET-30a-ompl9,expressed and purified Brucella protein omp19, and further validate its antigenicity by Western-blot.An indirect enzyme-linked immunosorbent assay was established using the purified recombinant protein omp19as coating antigen to detect antibody of B.Melitensis.Serum samples(n=180) from sheep were simultaneously tested by the iELISA and SAT,the data was analysised by SPSS.The data suggested that positive coincidence rate was96.81%,negative coincidence rate was77.91%, Kappa value was0.7531.The difference of these two detective methods was consistent,the diagnostic total coincidence rate could reach87.78%,the iELISA could be used as an alternative method for serological diagnosis of Brucellosis.The study two in order to find out the same and difference of B.abortus,B.melitensis and B.suis biological standard strains of type I including544A、16M、1330S and the corresponding vaccine strains S19、M5、S2, to explore the possibilities as a control strains when used to identify Brucella. We use eight kinds of ways to compare the six strains of bacteria including the biological classification,16SrDNA,BCSP31-PCR,AMOS-PCR,Mul tiple PCR,MLVA,MLST and PFGE. The result show that the similarity of544A and S19,1330S and S2are very high,but16M and M5has differences.The biological classification,16SrDNA,BCSP31-PCR,AMOS-PCR show that three of them could be replaced;1330S and544A could be replaced by S2and S19in the ways of multiple PCR and PFGE;1330S also could be replaced by S2in MLST.MLVA could tell the difference between the vaccine strains and the standard strains.We got a conclusion that S19and S2could be instead of544A and1330S as reference strains,but M5and16M only in some experiments could. All of them are not used in toxicity experiment.The study three in order to understand the prevalence,the KAP and cognition of Brucellosis in the siziwangqi in Inner Mongolia,we chose the farming and pastoral as study,we selected five villages from each region and extracted30household,the household survey was used to χ2test,the serum samples were used to laboratory examination.In this survey we found that the positive prevalence was16.67%,the incidence of pastoral was11.7%,the agricultural areas was7.90.%.The local villagers had lower cognitive rate on Brucellosis.so the risk factor increased,we should try the best to popularize Brucellosis knowledge.focusing on personal protection.