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

发布时间：2018-03-07 01:12

本文选题：超广谱β-内酰胺酶基因　切入点：实时荧光定量PCR　出处：《中国家禽》2017年06期 　论文类型：期刊论文

【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。
[Abstract]:In order to monitor bacterial drug resistance genes in commercial broiler farms, 74 strains of Enterobacteriaceae were isolated from commercial broiler farms, including 36 strains of Escherichia coli and 12 strains of Klebsiella spp. Fourteen kinds of 尾 -lactam antimicrobial agents were selected for drug sensitivity test. The drug resistance rates of Escherichia coli, Klebsiella and Proteus to various drugs were 26.922.The drug resistance rates of Escherichia coli, Klebsiella and Proteus to various drugs were 26.92% and 6.67%, 57.69% and 63.97%, respectively, and the phenotypes of extended-spectrum 尾 -lactamase ESBL were determined. The results showed that the three genotypes of Escherichia coli, Klebsiella sp. 96.51.7and Proteus were detected by PCR, and the detection rates of three genotypes in ESBL producing strains were 100 93.59.59.69.39.39.39.30.The detection rates of three genotypes in Escherichia coli, Klebsiella klebsii and Proteus were the highest, respectively, and the positive rates of the three genotypes were 79.39.3.The results showed that the three genotypes of Escherichia coli, Klebsiella klebsii, and Proteus were the highest. The real-time detection of drug resistance gene was carried out as dominant gene. The blaTEM gene was detected by real-time fluorescence quantitative PCR in three stages of four broilers' farm empty period and breeding stage, the results showed that: 1. The increase of blaTEM in henhouse with long breeding time and poor management was increased.
【作者单位】：山东畜牧兽医职业学院;安徽农业大学动物科技学院;
【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)
【分类号】：S852.61

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