基于高通量测序的一株鸭疫里氏杆菌的耐药表型和耐药基因分析

Analysis of the drug resistance phenotype and drug resistance genes of a Riemerella anatipestifer isolate by high-throughput sequencing

为研究鸭疫里氏杆菌耐药表型与耐药基因的关系,从浙江某鸭场病鸭体内分离到1株病原菌,经菌落形态和革兰染色特性观察、生化鉴定、PCR鉴定、动物回归试验,确定其为鸭疫里氏杆菌强毒株,血清凝集试验表明其为血清11型;药物敏感性分析结果显示,该菌对多黏菌素、复方新诺明、庆大霉素、四环素等耐药,对氨苄西林、头孢拉定、头孢唑林等敏感,属于多重耐药菌株;通过基因组高通量测序分析,检测到与表型相关的多黏菌素、红霉素、四环素等相关耐药基因,同时检测到多药外排泵及多药耐药蛋白基因,耐药表型与耐药基因检测结果相符。结果表明,通过高通量测序技术能准确获得鸭疫里氏杆菌分离株抗生素耐药的相关基因信息,对鸭疫里氏杆菌病的临床用药具有一定指导意义。

To study the relationship between resistance phenotype and resistance gene of Riemerella anatipestifer（ RA）, one pathogenic bacterium was isolated from diseased ducks in one duck farm in Zhejiang Province.The isolate was confirmed as a virulent strain after colonial morphology identification,biochemical identification,PCR identification and animal regression experiment.The serum agglutination test further showed the isolate was the RA 11 serotype,named as RA11 ZJ.Drug susceptibility experiments showed the isolated strain was a multidrug-resistant strain.The isolate was resistant to polymyxin,cotrimoxazole,gentamicin,tetracycline,and was sensitive to ampicillin,cefradine,and cefazoline.RA11 ZJ was sequenced on the Illumina Miseq platform.Polymyxin,erythromycin,tetracycline and other related drug-resistance genes were detected.The results also showed RA11 ZJ had multidrug efflux pumps and multidrug resistance protein genes.The results of drug resistance phenotype and drug resistance genes were consistent.The information of drug-resistant genes of RA11 ZJ was provided through high-throughput sequencing,it also provided important data sup port for clinical antibacterial therapy.