牦牛肉中阿维菌素残留的时间分辨荧光免疫层析检测方法的建立  

Establishment of Time-Resolved Fluorescence Immunochromatographic Method for Detection of Abamectin Residues in Yak Meat

作　　者：张正英[1] 崔乃元 高海燕 刘海珍[1] 黄文颖[1] 郝云晴 马立才 ZHANG Zhengying;CUI Naiyuan;GAO Haiyan;LIU Haizhen;HUANG Wenying;HAO Yunqing;MA Licai(Qinghai Provincial Animal Disease Prevention and Control Center,Xining 810001,China;Beijing WDWK Biotechnology Co.Ltd.,Beijing 100095,China;Inner Mongolia Autonomous Region Product Quality Inspection and Research Institute,Hohhot 010020,China)

机构地区：[1]青海省动物疫病预防控制中心,青海西宁810001 [2]北京维德维康生物技术有限公司,北京100095 [3]内蒙古自治区产品质量检验研究院,内蒙古呼和浩特010020

出　　处：《肉类研究》2020年第8期57-62,共6页Meat Research

基　　金：青海省科技计划基础研究计划项目(2018-ZJ-767)。

摘　　要：为建立定量检测牦牛肉中阿维菌素残留的时间分辨荧光免疫层析方法,以羧基化铕微球作为荧光标记物,与阿维菌素鼠单克隆抗体G10703进行共价偶联后喷涂于释放垫上,将阿维菌素抗原和羊抗鼠免疫球蛋白G分别包被于硝酸纤维素膜上作为检测线(T)和质控线(C),制备免疫层析试纸条;利用牦牛肉样本阿维菌素添加量和对应的T线与C线荧光信号峰值比值(T/C)拟合得到四参数标准曲线,建立牦牛肉中阿维菌素残留的时间分辨荧光免疫层析检测方法,并对所建立方法的灵敏度、准确度、特异性、精密度和稳定性等进行评价,用液相色谱-串联质谱方法对所建立方法的性能进行确证。结果表明:本方法对牦牛肉中阿维菌素的定量限为1.0μg/kg,加标回收率为86.9%~105.2%,与阿维菌素、伊维菌素、多拉菌素及依普菌素的交叉反应率分别为100.0%、73.1%、42.6%和97.5%,批内变异系数为5.4%~9.5%,批间变异系数为7.5%~11.2%,该方法准确度、精密度、灵敏度均较高,且性能稳定。The purpose of this study was to establish and evaluate a time-resolved fluorescence immunochromatographic method for quantitative detection of abamectin residues in yak meat.Anti-avermectin mouse monoclonal antibody G10703 was covalently coupled to carboxylated europium microspheres as a fluorescent marker,which were then sprayed on a release pad,and avermectin antigen and goat anti-mouse antibody were coated on nitrocellulose membrane as the detection line(T)and quality control line(C),respectively to make an immunochromatographic test strip.Finally,a time-resolved fluorescence immunochromatographic method for the detection of abamectin residue in yak meat was established by fitting the four-parameter standard curve between the amount of abamectin added to yak meat samples and the fluorescence signal peak ratio between T-line and C-line(T/C),and its sensitivity,stability,specificity,accuracy and precision were evaluated and compared with those of liquid chromatography-tandem mass spectrometry(LC-MS/MS).The limit of quantification of this method was 1.0μg/kg,and the spiked recovery was 86.9%–105.2%.The rates of cross-reactivity with avermectin,ivermectin,doramectin and iprodione were 100.0%,73.1%,42.6%and 97.5%,respectively.The within-batch coefficient of variation was 5.4%–9.5%,and the between-batch coefficient of variation was 7.5%–11.2%.The method had high accuracy,precision,sensitivity and stable performance.

关 键 词：牦牛肉 阿维菌素 时间分辨荧光免疫层析 

分 类 号：TS251.5[轻工技术与工程—农产品加工及贮藏工程]