Brucella Pulsed-field Gel Electrophoresis Typing

Objective To analyze the Polymorphism of the different strains of Brucella in our country in different times and atypical strain separated from our country, to develop the proper endonuclease map and experiment method, to explore the research on the type of Brucella. Put the PFGE data of Brucella into the web data PulseNet China, which will be made as the basic data on Brucellain China, to set up the basement of the web research on the molecule biology of Brucella, to provide database on the molecular biological research on the molecular epidemic of Brucellosis.Methods Set up the PFGE experiment method suitable for Brucella according to the experiments on the strains in the world, then we choose endonuclease XbaI. After many experiments on the referenced trains, we validated the repeatability of this experiment, and make the conditions of the experiment stable. Further, we detect the 122 strains separated from our country.Results Analyzing the XbaI PFGE map of the referenced strains, we found strains from the bovine, ovine and type1,3 of porcine are stable strains, and the others show weak similarity, which was according with the classification history on the Brucella. That means the strains from canine is the transfer host of the strains from the bovine; the other strains are the wild type cycled in and conserved by the nature. So we proved the objectivity of the PFGE method. We provided the molecular biology data for the classification on Brucella. And we found the distribution character of the nuclease sequence of the different species and biological strains Conclusions Considering the PFGE detecting result of 122 strains separated form our country, we found the most strains can get the good data, even can give a explain for the newly happened epidemic, and can apply in the research on the molecular epidemic character analysis, but there were some strains differ from the classical detect method. The reason may involve in the improper original identification, variance of the strains, and the mobile genome, which we can not make a conclusion now. We can say from our experiment result, the most important thing of our work is to find and set up a genome detecting method on Brucella. At the same time, we built the PulseNet China web database on Brucella in our country. While further improve this data, to enrich it, we hope it will benefit for the research and prevention and control of Brucellosis.