果蔬中15種農藥殘留快速檢測膠體金免疫層析多聯卡的研制

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

我國各大農貿市場的果蔬農殘監測以快速檢測為主，其中膠體金免疫層析技術（GoldImmuneChromatographicAssay，GICA）因其便捷性成為主流方法[1-5]。由于抗原與抗體一一對應特性，單個抗體的檢測卡僅能檢測一種目標物，難以滿足多種特定農藥殘留同時篩查檢測的需求，而個性化定制的多聯卡則可解決這個問題[6-7]。目前，可同時檢測果蔬中多種農殘的多聯卡技術報道較少[8]。本文基于競爭性GICA法，研制出15種常見農藥殘留（克百威、滅多威、丙溴磷、甲基異柳磷、辛硫磷、多菌靈、吡蟲啉、吡唑醚菌酯、甲霜靈、噻蟲嗪、氯蟲苯甲酰胺、水胺硫磷、百草枯、三唑磷和蠅毒磷）同步快速檢測多聯卡，并對檢測卡的主要性能指標進行評價，旨在為我國蔬果中多農藥殘留的個性化、高通量快速檢測以及食品安全提供科技支撐和技術保障。

1材料與方法

1.1材料與試劑

水果、蔬菜樣品，購于當地農貿市場。

農藥標準品，壇墨質檢科技股份有限公司；抗體及抗原，江南大學；牛血清白蛋白、檸檬酸三鈉、氯金酸溶液，美國Sigma公司；羊抗鼠二抗，北京博奧龍免疫技術有限公司；聚乙二醇20000、聚乙烯吡咯烷酮、吐溫-20，上海Aladdin公司；硝酸纖維素膜，德國Sartorius公司。

1.2儀器與設備

UV-2550紫外-可見光分光光度計（日本島津公司）；單通道及多通道可調移液器（德國Eppendorf公司）；超純水儀（美國Pall公司）；Allegra64R臺式高速離心機（美國Beckman公司）；SpectraMax@i3多功能酶標儀（美國MolcularDevices公司）；Mastersizer2000激光粒度儀（英國馬爾文儀器有限公司）；WEF-100噴金劃膜一體機（威爾芬）；WEF-200可編程自動切條機（威爾芬）；AMYQ-MR智能讀卡儀（傲敏生物）。……