胶体金免疫层析法高通量测定动物肌肉组织中多兽药残留  

High-throughput determination of multiple veterinary drug residues in animal muscle tissues by colloidal gold immunochromatographic assay

作　　者：贾先春 周嘉明 智军海 郝秋艳 杨晓芳 王玮[1] JIA Xianchun;ZHOU Jiaming;ZHI Junhai;HAO Qiuyan;YANG Xiaofang;WANG Wei(College of Food Science and Technology/State Key Laboratory of Meat Quality Control and Cultured Meat Development/National Center of Meat Quality and Safety Control/Inspection and Testing Center for Quality of Meat-products(Nanjing),Ministry of Agriculture and Rural Affairs,Nanjing Agricultural University,Nanjing 210095,China;Centre Testing International Group Co.Ltd.,Beijing 101102,China;Agricultural Products Quality and Safety Inspection and Testing Center,Kizilsu Kirgiz Autonomous Prefecture,Xinjiang Uygur Autonomous Region,Kizilsu Kirgiz Autonomous Prefecture 845350,China)

机构地区：[1]南京农业大学食品科学技术学院/肉品质量控制与新资源创制全国重点实验室/国家肉品质量安全控制工程技术研究中心/农业农村部肉及肉制品质量检验测试中心(南京),江苏南京210095 [2]华测检测认证集团股份有限公司,北京101102 [3]新疆维吾尔自治区克孜勒苏柯尔克孜自治州农产品质量安全检验检测中心,新疆维吾尔自治区克孜勒苏柯尔克孜自治州845350

出　　处：《南京农业大学学报》2024年第5期967-978,共12页Journal of Nanjing Agricultural University

基　　金：国家重点研发计划项目(2022YFF1100800)。

摘　　要：[目的]本文旨在建立一种胶体金免疫层析技术高通量测定猪肉、鸡肉组织中8种磺胺类、3种四环素类、2种喹诺酮类和甲氧苄啶药物残留的方法。[方法]动物肌肉组织中磺胺类、四环素类、喹诺酮类和甲氧苄啶经磷酸缓冲液提取,并经含Tween 20的磷酸缓冲液稀释后,与胶体金标记的单克隆抗体结合,抑制抗体与硝酸纤维素膜检测线(T线)上的抗原结合,从而导致T线颜色变化。通过比较T线和质控线(C线)颜色深浅,对试样中多兽药残留进行定性判定。[结果]本试验优化了pH值、抗体添加量、抗原包被量、Tween 20添加量和冻干保护剂种类等关键因素,磺胺类、四环素类、喹诺酮类和甲氧苄啶等药物的检出限分别为50、100、50、35μg·kg^(-1),灵敏度≥97%,假阳性率≤2%,假阴性率≤3%,且与现有标准方法检测结果一致。[结论]该方法操作简便,灵敏度和准确度高,检测时间短且稳定性好,可用于动物肌肉组织中多兽药残留的高通量快速检测。[Objectives]The purpose of this paper was to develop a method for the high-throughput determination of 8 sulfonamides,3 tetracyclines,2 quinolones and trimethoprim drug residues in pork and chicken tissues by colloidal gold immunochromatographic assay(GICA).[Methods]Sulfonamides,tetracyclines,quinolones and trimethoprim in animal muscle tissues were extracted by phosphate buffer and diluted by phosphate buffer with Tween 20,binding to colloidal gold-labeled monoclonal antibodies and inhibiting the binding of the antibodies to the antigen on the nitrocellulose membrane test line(T-line),which resulted in a change of the color of the T-line.The qualitative determination of poly-veterinary drug residues in the samples was made by comparing the color shades of the T-line and control line(C-line).[Results]This test optimized the key factors such as pH value,antibody addition,antigen encapsulation,Tween 20 addition and type of lyophilized protectant.The limit of detection for sulfonamides,tetracyclines,quinolones and trimethoprim was 50,100,50 and 35μg·kg^(-1),respectively,and the sensitivity≥97%,the false positive rate≤2%,the false negative rate≤3%,and the method was consistent with the results detected by the existing standard method.[Conclusions]The method was simple with high sensitivity,high precision,short detection time and good stability.It could be used for high-throughput rapid detection of multi-veterinary drug residues in animal muscle tissue.

关 键 词：多兽药残留 动物肌肉组织 胶体金免疫层析法(GICA) 高通量 检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]