基于反向荧光增强双信号通道免疫层析方法检测动物源食品中克百威残留  

Visualization-reverse Fluorescence Enhancement Immunochromatography(V-rFICTs)for the Determination of Carbofuran in Animal-Derived food

作　　者：孙晨晨 陈浩[3] 罗磊 胡荣荣 李盼 王玉龙[2] 刘鹏琰 杨剑婷[1] 张存政 SUN Chenchen;CHENG Hao;LUO Lei;HU Rongrong;LI Pan;WANG Yulong;LIU Pengyan;YANG Jianting;ZHANG Cunzheng(Anhui Science and Technology University,Fengyang 233100;Jiangsu Academy of Agricultural Sciences,Nanjing 210014;Taixing Agro-Tech Extension and Service Center,Taixing 225400;College of Food Science&Technology Nanchang University,Nanchang 330031)

机构地区：[1]安徽科技学院,安徽凤阳233100 [2]江苏省农业科学院,江苏南京210014 [3]泰兴市农业技术推广中心,江苏泰兴225400 [4]南昌大学食品学院,江西南昌330031

出　　处：《分析科学学报》2023年第6期656-664,共9页Journal of Analytical Science

基　　金：国家重点研发计划(2019YFC1605901)。

摘　　要：本研究建立了可视化-反向荧光增强双信号通道免疫层析技术(V-rFICTs),通过观察多巴胺包金的显色减弱以及量子点荧光的反向增强,实现对克百威的双信号、半定量检测。通过设计合成半抗原制备获得特异性单克隆抗体,以多巴胺包金核壳纳米材料标记抗体作为检测探针,将量子点(QDs-OVA)与包被原(CAR-OVA)喷涂于硝酸纤维膜(NC膜)上作为检测线(T线),同时喷涂兔抗鼠IgG抗体作为质控线(C线),组装成免疫层析试纸条。当样品不含克百威时,标记抗体结合在T线,裸眼可见明显紫红色条带,荧光猝灭,当克百威存在时,T线紫红色条带变弱或消失,而紫外下绿色荧光显现并随着克百威浓度的升高而增强。方法研究结果显示,V-rFICTs在可见光检测模式下的检测限为50 ng/mL,在荧光模式下的检测限为3.13 ng/mL,灵敏度提升15倍以上,15 min内可完成检测,与其他结构类似物无交叉反应。经方法验证,检测结果与液相色谱-串联质谱方法一致,具有较好的应用潜力。Carbofuran has been banned for use world-widely,due to its high toxicity and deadly threat to wild animals.Therefore,its residue in animal-derived food raised public concern.In this paper,we developed a visualization-reverse fluorescence enhancement immunochromatography strips(V-rFICTs)for the determination of carbofuran in animal-drive food by observing the decrease of purplish red bands of Au@PDA,and the increase of green fluorescence bands of quantum dots(QDs)on the T line,simultaneously.A hapten with carbofuran-like structure has been designed and synthesized,further induced a mAb with high specificity against carbofuran.Au@PDA nanoparticle was synthesized and conjugated with antibodies as test probe.V-rFICTs was fabricated for the determination of carbofuran.The results revealed that in the model of the colloidal gold-based immunochromatographic assay,the limit of detection(LOD)was 50 ng/mL,but reached 3.13 ng/mL under the condition of fluorescence readout signal.The sensitivity was promoted more than 15 fold.The detection could be finished within 15 minutes,with negligible cross-reactivity to the other analogues of carbofuran.Method validation was conducted by spiked chicken meat,which showed good consistency with the result of LC-MS/MS.The developed approach has great potential to be commercialized for use.

关 键 词：克百威 单克隆抗体 可视化-反向荧光增强双通道信号免疫层析法 动物源食品 

分 类 号：O658.1[理学—分析化学]