Development of A Multiplex Colloidal Gold Immunochromatographic Multicard for Rapid Detection of 15 Pesticide Residues in Fruits and Vegetables

NI Weihong'， YIN Danhan’，LI Gaotian1， XIAO Hailong1，LU Huajin²， SHEN Guofang' （1.Hangzhou Institute for Food and Drug Control Science，Hangzhou 31oo18， China; 2.Hangzhou Aomin Biotechnology Co.， Ltd.， Hangzhou 310019， China）

Abstract： In order to meet the needs of rapid detection of a variety of specific pesticide residues in fruits and vegetables，based oncoloidal gold immunochromatography technology，this paper developed acolloidal gold immunochromatographic multicard for the simultaneous rapid detection of 15 pesticide residues，such ascarbofuran， methocarb and profenofos，by optimizing the key factors such as coloidal gold-monoclonal antibody conjugate concentration and artificial antigen coating concentration.The test results showed that the detection limit of 15 pesticide residues was 0.01mg⋅kg^-1 to 0.3mg⋅kg^-1 ， no 1 negatives were found， the 1 positive rate was less than （20 5% ，and the specificity and accuracy were greater than 95% .A total of2 516 batches of samples were tested， and 6 batches of positive samples were detected.This method has good accuracy and simple operation，and is suitable for high-throughput rapid screening and detection of pesticide residues in vegetables and fruits.

Keywords： fruits and vegetables; pesticide residues; rapid detection; coloidal gold immunochromatographic multicard

我国各大农贸市场的果蔬农残监测以快速检测为主，其中胶体金免疫层析技术（GoldImmuneChromatographicAssay，GICA）因其便捷性成为主流方法[1-5]。由于抗原与抗体一一对应特性，单个抗体的检测卡仅能检测一种目标物，难以满足多种特定农药残留同时筛查检测的需求，而个性化定制的多联卡则可解决这个问题[6-7]。目前，可同时检测果蔬中多种农残的多联卡技术报道较少[8]。本文基于竞争性GICA法，研制出15种常见农药残留（克百威、灭多威、丙溴磷、甲基异柳磷、辛硫磷、多菌灵、吡虫啉、吡唑醚菌酯、甲霜灵、噻虫嗪、氯虫苯甲酰胺、水胺硫磷、百草枯、三唑磷和蝇毒磷）同步快速检测多联卡，并对检测卡的主要性能指标进行评价，旨在为我国蔬果中多农药残留的个性化、高通量快速检测以及食品安全提供科技支撑和技术保障。

1材料与方法

1.1材料与试剂

水果、蔬菜样品，购于当地农贸市场。

农药标准品，坛墨质检科技股份有限公司；抗体及抗原，江南大学；牛血清白蛋白、柠檬酸三钠、氯金酸溶液，美国Sigma公司；羊抗鼠二抗，北京博奥龙免疫技术有限公司；聚乙二醇20000、聚乙烯吡咯烷酮、吐温-20，上海Aladdin公司；硝酸纤维素膜，德国Sartorius公司。

1.2仪器与设备

UV-2550紫外-可见光分光光度计（日本岛津公司）；单通道及多通道可调移液器（德国Eppendorf公司）；超纯水仪（美国Pall公司）；Allegra64R台式高速离心机（美国Beckman公司）；SpectraMax@i3多功能酶标仪（美国MolcularDevices公司）；Mastersizer2000激光粒度仪（英国马尔文仪器有限公司）；WEF-100喷金划膜一体机（威尔芬）；WEF-200可编程自动切条机（威尔芬）；AMYQ-MR智能读卡仪（傲敏生物）。……