商品肉鸡场细菌超广谱β-内酰胺酶基因的荧光定量PCR检测  

Detection of Extended-spectrum β-lactamase Genes in Bacteria from Commercial Broiler Farm by Real-time PCR

作　　者：沈美艳 于海[2] 袁东芳 孙秋艳 李舫 

机构地区：[1]山东畜牧兽医职业学院,山东潍坊261061 [2]安徽农业大学动物科技学院,安徽合肥230036

出　　处：《中国家禽》2017年第6期20-23,共4页China Poultry

基　　金：山东省现代农业产业技术体系家禽创新团队建设项目(SDAIT-11-05)

摘　　要：为了对商品肉鸡场细菌耐药基因进行监控,从商品肉鸡场中分离肠杆菌科细菌74株,包括36株大肠杆菌、12株克雷伯氏菌、26株变形杆菌;选用14种β-内酰胺类抗菌药进行药物敏感性试验,检测分离菌株大肠杆菌、克雷伯氏菌和变形杆菌对各种药物的耐药率分别为26.92%~69.23%、8.67%~57.69%和63.97%~95.32%;经过超广谱β-内酰胺酶(ESBL)表型测定,确定19株耐药菌能够产生ESBL;用PCR对19株产生ESBL的菌株进行blaTEM、blaSHV和blaCTX-M三种基因型检测,显示大肠杆菌、克雷伯氏菌和变形杆菌中三个基因型检出率分别为100%、93.1%、96.5%;100%、51.7%、68.9%;100%、89.6%、79.3%。blaTEM检测检出率均为最高,作为优势基因进行耐药基因实时检测。利用实时荧光定量PCR对4个肉鸡场空舍期和养殖期三个时期地面样品进行blaTEM基因检测,结果表明,养殖时间较长和管理较差的鸡舍blaTEM增加倍数升高。To monitor the drug resistance in commercial broiler farms,74 Enterobacteriaceae strains,including 36 strains of E. coli, 12 strains of Klebsiella and 26 strains of Proteus were isolated and identified from broiler farms. 14 kinds of antibiotics were used to detect the drug resistance of those bacteria, and the resistance rate of the above bacteria were 26.92% to 69.23%,8.67% to 57.69% and 63.97% to 95.32% respectively. Through phenotype test,there were 19 strains of bacteria produced extended-spectrum β-1actamase （ESBL）, and three drug resistance genes, including blaTEM, blaSHV and blaCTX-M were detected in these bacteria. Positive rate of blaTEM, blaSHV, blaCTX-M in E. coli,Klebsiella and Proteus were 100%, 93.1%, 96.5% ; 100%, 51.7%, 68.9% ; 100%, 89.6%, 79.3% respectively. Gene of blaTEM had the highest positive rate and was used as the protogene and indicator gene. Real-time PCR was conducted to detect the blaTEM gene in the samples collected from ground at different breeding stages in broiler farms. The result indicated that good managing farm had lower increasing times of drug-resistance gene than the farms with bad management,and longer breeding period led to more increasing rate of drug-resistance rate.

关 键 词：超广谱β-内酰胺酶基因 实时荧光定量PCR 肉鸡场地面样品 

分 类 号：S855.1[农业科学—临床兽医学]