同时检测猪肉中4种抗菌药物残留的ELISA方法研究  

Simultaneous detection of four antimicrobial agents in pork by ELISA

作　　者：邢维维 荣佳 彭庆娟 秦誉 马立才 XING Weiwei;RONG Jia;PENG Qingjuan;QIN Yu;MA Licai(Beijing WDWK Biotechnology Co.Ltd.,Beijing 100095,China;New Hope Liuhe Co.,Ltd.,Beijing 100102,China)

机构地区：[1]北京维德维康生物技术有限公司,北京100095 [2]新希望六和股份有限公司,北京100102

出　　处：《畜牧与兽医》2022年第11期57-64,共8页Animal Husbandry & Veterinary Medicine

摘　　要：旨在建立一种前处理简单,且能同时检测磺胺甲噁唑、氟苯尼考、替米考星和甲氧苄啶的定性ELISA方法。磺胺甲噁唑、氟苯尼考、替米考星和甲氧苄啶抗原分别包被于不同的酶标微孔内,将待测样本与4种药物的单克隆抗体及酶标二抗充分混合,室温预反应1 min,取100μL上述混合反应液分别加至4种酶标微孔内进行检测。本研究通过对混合抗体稀释液及酶标二抗稀释液的配方进行筛选,建立了能同时检测猪肉中磺胺甲噁唑、氟苯尼考、替米考星和甲氧苄啶的ELISA方法,并对所建方法进行了敏感性、特异性、重复性等参数进行了评价。结果表明:本研究所建立ELISA方法对猪肉中磺胺甲噁唑、氟苯尼考、替米考星和甲氧苄啶的检出限为分别为25、10、15及10μg·kg^(-1),日内变异系数为2.52%~3.78%,日间变异系数小于5%。本研究建立的快速ELISA检测方法,具有前处理简单、检测时间短、结果可靠、重复性高等优点,为猪肉中相应抗菌药物残留的快速检测提供了可靠的新方法。This study was to establish a qualitative ELISA method for simultaneous detection of sulfamethoxazole,florfenicol,tilmicosin and trimethoprim.The sulfamethoxazole,florfenicol,tilmicosin and trimethoprim antigens were coated in different enzyme-labeled micro-wells.The samples were fully mixed with the monoclonal antibody and enzyme-labeled second antibody of the four drugs,and were incubated at room temperature for 1 min;100μL of the mixed solution was added to the above four kinds of micro-wells for detection.In this study,a new ELISA method was developed for simultaneous determination of sulfamethoxazole,florfenicol,tilmicosin and trimethoprim in pork by screening the formula of the mixed antibody diluent and the enzyme-labelled second anti-diluent;and the sensitivity,specificity and repeatability of the proposed method were evaluated.The results were that the detection limits of sulfamethoxazole,florfenicol,tilmicosin and trimethoprim in pork by ELISA were 25,10,15 and 10μg·kg^(-1),respectively,that the intra-day coefficient of variation was 2.52%-3.78%,and that the inter-day coefficient of variation was less than 5%.The rapid EISA established here possessed the advantages of simple pretreatment,short detection time,reliable results and high repeatability.This method provided a new and reliable method for rapid detection of antimicrobial drug residues in pork.

关 键 词：药残 多因子 ELISA 猪肉 

分 类 号：S859.84[农业科学—临床兽医学]