The Study Of MLVA Molecular Epidemiological Characteristics And Drug Sensitization In Vitro Of Brucella

Objects:The data of brucella biotype and MLVA classification in China,provinces and municipalities were collected and analyzed.The characteristics of the distribution were analyzed at different stages of brucellosis.The biotype and MLVA genotyping of Brucella strains isolates from Qinghai Province were analyzed.It was compared with the national Brucella strains.The establishment of the national brucella biotype and MLVA classification of the database for the application of molecular epidemiology to prevent brucellosis to provide a scientific basis.To compare the advantages and disadvantages of MH(Mueller-HintonA)agar plate,MH(Mueller-HintonA)agar blood plate and Brucella-Agar plate culture medium to make Brucella susceptibility test,the most suitable brucella drug sensitivity Test medium.180 strains of Brucella were selected for susceptibility screening.Rifampicin(RD),levofloxacin(LEV),moxifloxacin(MXF),streptomycin(S),cotrimoxazole(SXT),ceftriaxone sodium(CRO),amoxicillin/clavulanic acid(AMC)And tetracycline(DOS)8 commonly used in the first-line treatment of brucellosis patients and new antibiotics listed to explore the resistance of brucella.To analyze the difference of antimicrobial susceptibility of brucella isolated from high epidemic area and sporadic area in our country,so as to guide the treatment of brucellosis in clinical treatment and rational use.Methods:The combination of "brucellosis/brucella","biotyping,identification" and"multiple site number variable tandem repeat sequence analysis/MLVA" keyword combination in NCBI(National Biotechnology Information Center)Site,Wanfang database,Chinese knowledge network,Chinese periodical database(Vip)and other databases in recent years on the brucella MLVA classification of the core journals,Data on Brucella MLVA typing in Chinese provinces,autonomous regions and municipalities.The number of Brucella strains,the biological type,the separation host,the time,the local and the 16 VNTR site repeat results of MLVA panel 1 and panel 2 were analyzed and the number of samples was statistically analyzed for each experiment and literature.According to the American Clinical Laboratory Standards Institute CLSI/NCCLS 1999-1995 issued by the drug sensitivity test standards,sing K-B drug sensitive paper method,Rifampicin(RD),levofloxacin(LEV),moxifloxacin(MXF),streptomycin(S),cotrimoxazole(SXT),ceftriaxone sodium(CRO),amoxicillin/clavulanic acid(AMC)and tetracycline(DOS)and other eight kinds of antibiotics for susceptibility testing。Observe the antibacterial drug paper with or without antibacterial ring,the amount of its diameter,with "mm" number of records。Results:1、A total of 131 articles of brucella biology and genotyping were retrieved.A total of 52 literatures on brucella biology and genotyping with specific data were screened.2、The biological patterns of 1643 strains of Brucella were collected from 31 provinces of autonomous regions and municipalities,and the data of 16 VNTR site repeat numbers of host,time,place and MLVA panel 1 and panel 2 were isolated.3、Brucella strains were classified according to the biotype,B.melitensis accounted for 73.82%,B.abortus accounted for 15.16%,B.suis 113 accounts for 6.88%,B.canis Accounted for 4.14%.B.melitensis biovar 1 accounts for 6.35%;B.melitensis biovar 2 accounts for 4.62%;B.melitensis biovar.3 accounting for 89.0%.4、The genotypes are mainly composed of 42,43,belong to the eastern Mediterranean for the MLVA-8 parting5、There are more genotypes of MLVA8 in Guangdong,Hainan,Fujianand Shanxi,Inner Mongolia,Xinjiang,Ningxia,and Qinghai provinces,where the epidemic of brucellosis is serious,42,43,63,114,58,112,47,45 and 96 genotypes,be showing the diversity of genotype..6、The Brucella were isolated from Qinghai Province,Brucella divided into three species(B.melitensis,B.abortus,B.suis)and 6 biotypes(B.melitensis biovar 1,2,and 3;B.abortus biovar 3and7;B.suis biovar 1).B.melitensis accounted for 91.38%,B.abortus accounted for 6.90%,B.suis accounted for 1.72%.B.melitensis biovar3 is the main dominant species.in Qinghai Province7、The brucella of Qinghai Province were divided into 3 populations 7 clusters(A～G group)40 genotypes,According to MLVA8 type there are 42,43,47,28,63,36,112,6,42 and 43 type is the main.8、MH(Mueller-Hinton)agar plates and MH(Mueller-Hinton)agar blood plates are not suitable for Brucella growth.9、Brucella has a common line of streptomycin,doxycycline,amoxicillin/clavulanic acid,levofloxacin,ceftriaxone,rifampicin and moxifloxacin Antibiotics are sensitive,no drug resistance phenomenon In the Brucella susceptibility test。10、39%of Brucella strains were resistant to cotrimoxazole,and the distribution of brucella-resistant Brucella strains in almost every province.Conclusions:1、Separation of the main Brucella biotype are not obvious features in the north and southern of China.There were isolated B.melitensis,B.abortus,and B.suis.B.canis is more isolated in the southern.2、MLVA 42 and 43 are predominantly in each province.MLVA8 genotype is more diverse,the genes were polymorphic distribution in the traditional brucellosis epidemic areas,the current epidemic of brucellosis epidemic provinces and the separation of Brucella strains more provinces such as Shanxi,Inner Mongolia,Xinjiang,Ningxia,Qinghai,Guangdong and Hainan.3、The existence of their own natural foci outbreak of the epidemic and foreign imports of internal and external transmission of the dual risk of attack in Qinghai.Bacterial disease prevention and control is extremely difficult.4、There are many of direct or indirect transmission as ground natural foci,livestock→people,wildlife→people,wildlife←→livestock→people，wildlife←→livestock,Input sexually transmitted.5、There is mainly B.melitensis,mainly for the B.melitensis biovar 3,but also B.abortus biovar 3and7 and B.suis biovar 1 in Qinghai.Brucella biological species was Diversity.6、Most of the B.melitensis strains of Qinghai are concentrated in a cluster.Interconversion between very little difference on it only one or two repeats difference,With other provinces,there are significant differences in panel2B,the difference between a plurality of repeat sequences.7、There are nine sharing genotype of Brucella strains in Qinghai with the genotype strains in Heilongjiang,Shanxi,Guangdong,Inner Mongolia,Hebei,Henan,Shaanxi,Jiangxi,Chongqing,Xinjiang and other provinces.8、Brucella-Agar is suitable for Brucella susceptibility testing.It be used as a culture medium for Brucella susceptibility experiments.9.Cotrimoxazole has a drug resistance phenomenon in the clinical medication of "Clinic disease diagnosis and treatment guidelines.10、25 provinces were tested for cotrimoxazole(SXT)resistant strains of Brucella.It is no difference in the distribution of north and south.Resistant to cotrimoxazole Brucella strains accounted for 39%(70/180).11、Suggested treatment of brucella patients should be combined with drug susceptibility test results research and observation results.