啶酰菌胺单克隆抗体制备及其试纸条的研究

Study on preparation of monoclonal antibody and test strip of boscalid

啶酰菌胺是一种新型烟酰胺类杀菌剂,常用来防治果蔬各种类型的真菌病害。为构建一种啶酰菌胺简便、准确的检测方法,该实验首先应用啶酰菌胺人工抗原免疫BALB/c小鼠,免疫后取小鼠脾脏细胞与骨髓瘤细胞进行融合,筛选得到可产生啶酰菌胺单克隆抗体的杂交瘤细胞株,随后建立了啶酰菌胺的胶体金免疫层析方法。结果表明,所制备的啶酰菌胺单克隆抗体特异性强,选取了蔬菜水果中可能与啶酰菌胺联合用药的17种药物进行特异性测定,均未产生交叉反应,灵敏度高,半数抑制浓度(IC_(50))值为1.03 ng/mL;建立的胶体金免疫层析法对蔬菜水果的检测限为5μg/L,符合GB 2763—2021《食品安全国家标准食品中农药最大残留限量》规定的最大残留限量要求,可用于实际样品中啶酰菌胺残留的快速检测,具有良好的应用前景。

Boscalid is a new kind of nicotinamide fungicides,which is used to control various kinds of fungal diseases of fruits and vegetables.In order to establish a simple and accurate method for boscalid detection,BALB/c mice were immunized with artificial antigen of boscalid,after the spleen cells of mouse were fused with myeloma cells to screen hybridoma cell lines that could produce monoclonal antibodies to boscalid.Then,a colloidal gold immunochromatography method for boscalid was established.The results showed that the prepared monoclonal antibody was highly specific.17 kinds of drugs in vegetables and fruits that might be used in combination with the boscalid were selected for specific determination.No cross-reaction occurred,and the sensitivity was high,and the half inhibitory concentration(IC_(50))value was 1.03 ng/ml.The detection limit of the established colloidal gold immunochromatography for vegetables and fruits was 5μg/L,which met the requirements of maximum residue limit specified in GB 2763—2021"National food safety standard-Maximum residue limits for pesticides",and could be used for the rapid detection of the boscalid residues in actual samples,which had a good application prospect.