非布鲁菌病疫区江苏省连云港市13株布鲁菌分离株的MLVA基因分型特征分析
MLVA genotyping characteristics of 13 strains of Brucella isolated from non-brucellosis epidemic area Lianyungang City of Jiangsu Province
摘要目的:了解非布鲁菌病疫区江苏省连云港市布鲁菌分离株的基因分型特征。方法:对2018年连云港市第一人民医院临床微生物室血培养分离的13株疑似布鲁菌菌株进行初步鉴定;同时,采用实时荧光定量PCR(Real-time PCR)检测布鲁菌特异性基因bcsp31和插入序列IS-711,对鉴定结果进行复核和分型。运用多位点可变数目串联重复序列分析(MLVA)进行基因分型,测序结果用Mega 4.0软件进行编辑。结果:13株分离株经初步鉴定均为布鲁菌属。经Real-time PCR检测,13株分离株均为羊种布鲁菌。MLVA结果显示,13株羊种布鲁菌分为12个基因型,且聚类在"中地中海簇"中。13株羊种布鲁菌中有3株生物1型、2株生物2型和8株生物3型。结论:连云港市分离的布鲁菌菌株均为羊种布鲁菌,MLVA聚类在"中地中海簇"中。
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abstractsObjective:To study the genotyping characteristics of Brucella strains isolated from Lianyungang City (non-brucellosis epidemic area) of Jiangsu Province. Methods:Preliminary identification of 13 suspected strains of Brucella isolated from blood culture in Clinical Microbiology Laboratory of the First People's Hospital of Lianyungang City in 2018 was conducted; at the same time, the specific gene bcsp31 and insertion sequence IS-711 of Brucella were detected by quantitative real-time PCR (Real-time PCR), and the identification results were rechecked and typed. Multiple locus variable-number tandem repeat analysis (MLVA) was applied for genotyping, and the sequencing results were edited by Mega 4.0 software. Results:All the 13 strains were identified as Brucella by preliminary identification. Real-time PCR confirmed that all the 13 strains were Brucella melitensis. The results of MLVA showed that 13 strains of Brucella melitensis were divided into 12 genotypes and clustered in the "middle Mediterranean cluster". Among 13 strains of Brucella melitensis, 3 strains were biovar 1, 2 strains were biovar 2 and 8 strains were biovar 3. Conclusion:All the Brucella strains isolated from Lianyungang City are Brucella melitensis and the MLVA cluster is in the "middle Mediterranean cluster".
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