Yang Xingdong, Qu Yang, Wang Chenchen, Wu Lihua, Hu Xiaofei
Institute of Food and Drug Inspection, College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou 466001, China.
Key Laboratory of Animal Immunology, Ministry of Agriculture and Rural Affairs & Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Foods. 2024 Dec 25;14(1):27. doi: 10.3390/foods14010027.
The ingestion of food contaminated with citrinin (CIT) poses a variety of health risks to humans and animals. The immunogens (CIT-COOH-BSA, CIT-H-BSA) and detection antigen (CIT-COOH-OVA, CIT-H-OVA) were synthesised using the active ester method (-COOH) and formaldehyde addition method (-H). A hybridoma cell line (3G5) that secretes anti-CIT monoclonal antibodies (mAbs) was screened via CIT-H-BSA immunisation of mice, cell fusion, and ELISA screening technology. The cell line was injected intraperitoneally to prepare ascites. The reaction conditions for the indirect competitive ELISA (-ELISA) were optimised, and an -ELISA method for detecting CIT was preliminarily established. The results revealed that the IC of CIT from optimised -ELISA was 37 pg/mL, the linear detection range was 5.9230 pg/mL, and the cross-reaction (CR) rate with other analogues was less than 0.01%. The intra-assay and interassay sample recovery rates of CIT were 84.792.0% and 83.6~91.6%, and the coefficients of variation (CVs) were less than 10%. The -ELISA of CIT established in this study was not significantly different from the HPLC results and is rapid, highly sensitive and strongly specific, providing technical support for the detection of CIT.
摄入被桔霉素（CIT）污染的食物会对人类和动物构成多种健康风险。采用活性酯法（-COOH）和甲醛加成法（-H）合成了免疫原（CIT-COOH-BSA、CIT-H-BSA）和检测抗原（CIT-COOH-OVA、CIT-H-OVA）。通过用CIT-H-BSA免疫小鼠、细胞融合和ELISA筛选技术，筛选出分泌抗CIT单克隆抗体（mAb）的杂交瘤细胞系（3G5）。将该细胞系腹腔注射制备腹水。优化了间接竞争ELISA（-ELISA）的反应条件，初步建立了检测CIT的-ELISA方法。结果表明，优化后的-ELISA对CIT的IC为37 pg/mL，线性检测范围为5.9230 pg/mL，与其他类似物的交叉反应（CR）率小于0.01%。CIT的批内和批间样品回收率分别为84.792.0%和83.6~91.6%，变异系数（CV）小于10%。本研究建立的CIT的-ELISA与HPLC结果无显著差异，具有快速、高灵敏度和强特异性的特点，为CIT的检测提供了技术支持。
