Establishment Of Diagnostic Methods And Epidemiological Investigation For Brucellosis And Lyme Disease

Objective: Firstly,to investigate the molecular prevalence of Borrelia and Brucella in parasitic and free-living ticks in several Cities/Counties of Xinjiang Province,and furt her to provide the theory basis for confirmation dominant ticks as host for these two bacteria;secondly,to carry out the etiology and serological investigation of the Lyme disease and brucellosis of the shepherds in Tacheng City,north region of Xinjiang Province,which provides basic information of the local impacts of these two diseases;and thirdly,to prepare and select the corresponding monoclonal antibody using the purified Borrelia burgdorferi Bmp A protein and Brucella Vir B5 protein for the later preparation of nanoparticles reagents.Methods: After the genomic DNA extraction was performed,the nucleotides of Borrelia and Brucella were detected from ticks collected in several Cities/Counties of Xinjiang Province.A total of 246 blood samples from the shepherds were used to detect the infectious rates for both pathogens by bacterial and serological methods.As for antigens of Borrelia burgdorferi Bmp A protein and Brucella Vir B5 protein,monoclonal antibodies were developed and screened after immunizing mice,cell fusion,cloning,outscreen of monoclonal cells,subtypes identification.Then the optimal antibodies for Bmp A and Vir B5 proteins were further screened with Western Blot methods.Results: The average positive rate for Brucella nucleotide was 7.73% in parasitic ticks.Among them,they were respectively 5.30%,8.74%,5.26% and 17.50% in Hyalomma asiaticum,Haemaphysalis punctata,Dermacentor marginatus and Rhipicephalus turanicus.The average positive rates for Borrelia were 3.86% and 35.07% in parasitic ticks and free-living ticks,respectively.Among them,they were respectively 5.30%,3.82% and 5.26% in Hy.Asiaticum,Ha.punctata and D.marginatus.The positive rates were 2.03% and 19.51% in 246 blood smples by bacterial and serological methods,respectively.The results of antibody titer test showed 12 cell strains for Bmp A protein and 6 cell strains for Vir B5 protein were positive by Western Blot.Two optimal cell strain for Bmp A protein and Vir B5 protein were further determined by Western Blot.Conclusion: Pasture ticks maybe play a certain role in the horizontal transmission of livestock brucellosis.Hy.asiaticum,Ha.punctate and D.marginatus become as Lyme disease carriers,which might expand the scope of Lyme disease vectors.The detection results of the blood samples in shepherds presented a high infection of Borrelia and Brucella,especially for Brucella,should strengthen the outscreen of blood donors in high prevalent area.The optimal cell strains for Bmp A protein and Vir B5 protein,which could secrete monoclonal antibodies,were obtained in this study.The findings would lay foundations for the later article preparation of nanoparticle reagent.