The Establishment And Application Of The Brucella MLVA And The HOOF Standard Operation Procedures

Objective:1.To establish the standard operating procedures on multiple locus variable number tandem repeats analysis typing and hypervariable octameric oligonucleotide finger-prints on brucella,as brucellosis epidemiological surveillance tool。2.the methods used to provide reference67brucella isolates form Inner Mongolia,1984-1995.Methods:1.Extract the bacterial nucleic acid,from the16Brucella standard strains and67of Inner Mongolia1983-1997.2.Design and synthesis of the primers of MLVA and HOOF genotyping methods.3.the standard strains and local strains PCR,MLVA panel1using the agarose gel electrophoresis, Distinguish strip sequencing, MLVA panel2and HOOF using STR.4.Calculation of tandem repeat number,organized into an Excel table,Combined with BioNumerics (Version5.0) software,MLVAand HOOF genotyping analysis of Brucella strains.5.Established MLVA and HOOF standardized method.Results:1.Established MLVA and HOOF typing standardized methods by Standard strains of Brucella.2.This method can diacriminate brucella Inner Mongolia local strains.3.MLVA and HOOF typing results of16standard strains and53of Inner Mongolia bacteria establishment of a database.Conclusion:1.Preliminary established MLVA and HOOF standardized method2.These methods can be used for distinguish Brucella species, trace the source of infection,determine the prevalence areas.