畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌的LAMP检测及其耐药性分析

doi:10.16431/j.cnki.1671-7236.2020.07.039

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (7): 2325-2335.doi: 10.16431/j.cnki.1671-7236.2020.07.039

• 环境安全 • 上一篇

畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌的LAMP检测及其耐药性分析

范丽霞^1,2, 苑学霞^1,2, 李鸳鸯¹, 邬元娟^1,2, 赵善仓^1,2, 张丙春^1,2, 王文博^1,2

1. 山东省农业科学院农业质量标准与检测技术研究所, 山东省食品质量与安全检测技术重点实验室, 济南 250100;
2. 山东省农业科学院-北卡罗莱纳州立大学农产品检测技术和风险评估联合实验室, 济南 250100

收稿日期:2020-01-21 出版日期:2020-07-20 发布日期:2020-07-18
通讯作者: 王文博(1974-),男,山东龙口人,研究员,研究方向:农产品质量安全与风险评估,E-mail:wangwb74@163.com E-mail:wangwb74@163.com
作者简介:范丽霞(1983-),女,黑龙江北安人,博士,助理研究员,研究方向:农产品质量安全与风险评估,E-mail:superdemeter@163.com
基金资助:
国家重点研发计划子课题（2016YFD05014072）；国家重点研发计划政府间国际科技创新合作重点专项（2016YFE0109200）；国家自然科学基金项目（41907030）

LAMP Detection and Drug Resistance Analysis to Staphylococcus aureus and Escherichia coli in Livestock and Poultry Manure

FAN Lixia^1,2, YUAN Xuexia^1,2, LI Yuanyang¹, WU Yuanjuan^1,2, ZHAO Shancang^1,2, ZHANG Bingchun^1,2, WANG Wenbo^1,2

1. Shandong Provincial Key Laboratory of Test Technology on Food Quality and Safety, Institute of Agricultural Standards and Testing Technology for Agri-Products, Shandong Academy of Agricultural Sciences, Jinan 250100, China;
2. NCSU-SAAS Joint Laboratory for Testing Technology & Risk Assessment for Agro-products, Jinan 250100, China

Received:2020-01-21 Online:2020-07-20 Published:2020-07-18

摘要/Abstract

摘要： 为深入了解畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌的流行及耐药状况，采用建立的畜禽粪便中金黄色葡萄球菌和大肠埃希氏菌的环介导等温扩增技术（LAMP）检测方法对山东省内牛粪污、猪粪、鸡粪、鸭粪中的金黄色葡萄球菌和大肠埃希氏菌进行检测，进而对检测阳性样品进行分离，并对药敏试验中多重耐药性菌株进行耐药基因预测。结果表明，LAMP方法检测畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌与国家标准方法检测结果符合率为100%，重复性好，特异性高。从80份畜禽粪污样品中分离到的3株金黄色葡萄球菌均对青霉素耐药；34株大肠埃希氏菌对氟苯尼考、利福平的耐药比例高于50%，高于25%以上的还有氨苄西林、四环素、多西环素、磺胺异噁唑、头孢噻吩和头孢噻呋；耐药基因预测结果显示，鸡粪、鸭粪、牛粪和猪粪中的4株大肠埃希氏菌分别携带10、8、20和15种耐药基因；鸡粪中的1株金黄色葡萄球菌携带11种耐药基因。说明山东地区畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌耐药状况严峻，菌株普遍多重耐药，且携带多种耐药基因。

关键词: 畜禽粪污; LAMP; 金黄色葡萄球菌; 大肠埃希氏菌; 药敏试验; 耐药基因

Abstract: For further understanding of the prevalence and drug resistance of Staphylococcus aureus and Escherichia coli in livestock and poultry manure,the established loop-mediated isothermal amplification technique (LAMP) in livestock and poultry manure was used to detect S.aureus and E.coli in cattle,pig,chicken and duck manure in Shandong province.Then the strains were isolated from the positive samples,and the resistance genes of the multiple resistance strains in the drug sensitivity test were predicted.The results showed that the LAMP method to detect S.aureus and E.coli in livestock and poultry manure was completely consistent with the national standard method,with good repeatability and high specificity.Three strains of S.aureus isolated from 80 samples of livestock and poultry manure were resistant to penicillin.The drug resistance rate of 34 E.coli strains to florfenicol and rifampicin was more than 50%,and the proportion of drug resistance to ampicillin,tetracycline,doxycycline,sulfisoxazole,cephalotin and ceftiofur were more than 25%.The prediction results of drug resistance gene showed that four strains of E.coli in chicken,duck,cow and pig manure carried 10,8,20 and 15 drug resistant genes,respectively.One strain of S.aureus in chicken manure carried 11 drug resistant genes.It indicated that the drug resistance situation of S.aureus and E.coli in livestock and poultry manure of Shandong was severe,the strains were generally multi-drug resistant,and they carried a variety of drug resistance genes.

Key words: livestock and poultry manure; LAMP; Staphylococcus aureus; Escherichia coli; drug sensitivity test; resistance gene

中图分类号:

S852.6

范丽霞, 苑学霞, 李鸳鸯, 邬元娟, 赵善仓, 张丙春, 王文博. 畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌的LAMP检测及其耐药性分析[J]. 中国畜牧兽医, 2020, 47(7): 2325-2335.

FAN Lixia, YUAN Xuexia, LI Yuanyang, WU Yuanjuan, ZHAO Shancang, ZHANG Bingchun, WANG Wenbo. LAMP Detection and Drug Resistance Analysis to Staphylococcus aureus and Escherichia coli in Livestock and Poultry Manure[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 2325-2335.

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畜禽粪污中金黄色葡萄球菌和大肠埃希氏菌的LAMP检测及其耐药性分析

LAMP Detection and Drug Resistance Analysis to Staphylococcus aureus and Escherichia coli in Livestock and Poultry Manure

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