【作者】 郭军庆； 杜振昆； 张妙仙； 项玉燕； 柯永恩； 周继勇；

【机构】 浙江大学动物预防医学研究所； 浙江大学传染病诊治国家重点实验室人兽共患病研究室； 浙江舟山市畜牧兽医局； 浙江金华市畜牧兽医局；

【摘要】 根据布氏杆菌编码36kDa外膜蛋白的omp2基因多态性设计引物,PCR扩增51株布氏杆菌分离株omp2a和omp2b基因,并进行序列测定,GenBank数据库BLAST序列比对分析显示,43株为牛种布氏杆菌(B.abortus),8株为羊种布氏杆菌(B.millitensis),未能区分布氏杆菌生物型。转座因子IS711在布氏杆菌不同种属中存在的多态性,利用IS711通用引物和种特异性引物对布氏杆菌分离进行AMOS-PCR检测,鉴定B.abortus 5、6、9或3b型35株,B.abortus 1、2或4型2株,B.melitensis 1或2型5株,另有9株未能鉴定生物型。PCR扩增未分型9株细菌的rpoB基因(4134 bp),并进行序列分析,结果鉴定B.melitensis 1型3株、B.abortus 1或4型1株,B.abortus 7型1株,其余4株无法确定其生物型。更多还原

【Abstract】 To explore molecular typing for Brucella isolates,we designed specific primers according to DNA polymorphism of omp2 encoding a 31 kDa.outer membrane protein.The omp2a and omp2b fragments of 51 Brucella isolates were amplified by PCR and sequenced.BLAST in GenBank database showed that 43 of 51 brucella isolates were B.abortus and 8 strains B. millitensis,but their biotypes failed to be identified.As the method of AMOS-PCR can detect the biotype of the brucella based on the species-specific IS711 gene,which is a transposable element and presents the insert position polymorphism in the difference biotypes of Brucella,the consensus primer and 5 specific primers in downstream were then designed for Brucella biotyping.The AMOS-PCR identified 35 strains as B.abortus 5,6,9 or 3b,2 strains of B.abortus 1,2 or 4,and 5 strains of B. melitensis.However,there were 9 strains not be clasSified into any biotypes.A rpoB gene of B.melitensis with a length of 4134 bp includes all the polymorphism of species and biotype.RpoB genes of the unidentified 9 strains were then amplified by PCR and sequenced for biotyping.Three strains of B.melitensis 1,1 strain of B.abortusl or 4,and 1 strain of B.abortus 7 were identified,whereas 4 strains of Brucella isolates failed to be identified the biotype of species yet.更多还原