作　　者： ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;

机构地区： 深圳出入境检验检疫局

出　　处： 《动物医学进展》 2013年第8期7-12,共6页

摘　　要： 近年来,由于抗生素滥用导致细菌产生耐药性的现象已经非常严重,而产超广谱β-内酰胺酶(ESBLs)大肠埃希菌耐药菌株的出现,已成为目前出入境食品安全检测的重点和难点。耐药大肠埃希菌ESBLs基因型有5种,其中产CTX-M和TEM两种基因型最为普遍。为了加强对产CTX-M和TEM两种基因型ESBLs耐药大肠埃希菌的检测,本研究在反复优化的基础上,建立了针对产CTX-M和TEM两种基因型ESBLs大肠埃希菌的快速检测方法。针对CTX-M基因型的PCR方法检测CTX-M ESBLs型的灵敏度为10-5,针对TEM基因型的PCR方法检测TEM ESBLs型的灵敏度为10-4,双重PCR方法检测对应基因型ESBLs灵敏度均为10-4。该方法特异性好,尚未发现假阴性和假阳性现象。采用建立的方法与VITEK-2细菌鉴定系统的鉴定方法和NCCLSM100-S10推荐的检测方法同时对623份样品进行检测并比较,结果表明,共检出只含CTX-M型ESBLs的耐药菌株13株,检出只含TEM基因型ESBLs的耐药菌株1株,同时含有CTX-M和TEM两种基因型ESBLs的耐药菌株9株,检测结果与细菌分离鉴定结果一致。表明所建立的方法可为产ESBLs大肠埃希菌的快速准确检测提供有效的工具,能够满足产ESBLs大肠埃希菌快速、准确、有效的口岸检疫需求。 Antimicrobial resistance became more and more serious with the abuse of antimicrobial agents in animals and humans. E. coli were easy to obtain multidrug-resistance, especially for extended-spectrum β- lactamases （ESBLs） and became prevalence in the animal products. ESBLs E. coli was a major problem of food safety for inspection and quarantine at port. CTX-M ~ TEM gene types of ESBLs have become domi- nant with much greater penetration into E. coli among five gene types of ESBL-producing organism. It was urgent to develop a rapid molecular to detect the ESBL-producing E. coli in entry-exit animal food. A PCR assay for dominant CTX-M TEM gene types of ESBLs was developed and evaluated with the comparison of VITEK-2 system and NCCLSM100-S10 standard recommend by clinical and laboratory standards insti- tute with 623 samples. 13 strains of CTX-M gene type of ESBLs,1 of TEM and 9 of both were tested. The specificity of assay developed was good without any cross reaction with other gene types of ESBL-produ- cing organism and other bacteria. The sensitivities of PCR assay was 10-s dilution of CTX-M standard E. coli and 10-4 dilution of TEM. The sensitivities of the duplex assay was 10-4 dilution for both genes. These methods potentially allowed for more rapid, sensitive, and specific detection of animal products of import and export.

关 键 词： 超广谱 内酰胺酶 耐药性 快速检测

领　　域： [农业科学] [农业科学] [农业科学]