高灵敏度呕吐毒素时间分辨荧光快速定量检测卡的研发和应用  

Development and Application of High Sensitivity Time-Resolved Fluorescent Rapid Quantitative Detection Card for Deoxynivalenol

作　　者：王振芳 王平东 邓庆 叶金 孙秀兰[4] 纪剑 谈婷 赵皖 肖理文 WANG Zhenfang;WANG Pingdong;DENG Qing;YE Jin;SUN Xiulan;JI Jian;TAN Ting;ZHAO Wan;XIAO Liwen(Central Grain Reserve Dezhou Direct Warehouse Co.,Ltd.,253000 Dezhou Shandong,China;Jiangsu Quality Inspection Center Co.,Ltd.,211155Nanjing,China;Institute of Grain and Oil Quality and Safety,National Food and Strategic Reserves Administration,102629 Beijing,China;School of Food Science,Jiangnan University,214122 Wuxi Jiangsu,China;Nanjing Micromeasurement Biotechnology Co.,Ltd.,210031 Nanjing,China)

机构地区：[1]中央储备粮德州直属库有限公司,山东德州253000 [2]中储粮江苏质检中心有限公司,南京211155 [3]国家粮食和物资储备局科学研究院粮油质量安全研究所,北京102629 [4]江南大学食品学院,江苏无锡214122 [5]南京微测生物科技有限公司,南京210031

出　　处：《粮食加工》2024年第1期102-107,共6页Grain Processing

摘　　要：利用时间分辨荧光快速定量检测技术采用稀有元素“铕”作为示踪物,开发出一种用于检测粮食谷物中的呕吐毒素含量的高灵敏度呕吐毒素时间分辨荧光快速定量检测卡。交联剂碳二亚胺(EDC)用量25 μL/mL、抗体量10 μg/mL标记脱氧雪腐镰刀菌烯醇DON单抗并用硼酸-硼砂缓冲液(12 mmol/L,pH 7.27)体系作为喷膜稀释液喷于结合垫上。Tris-HCl (10 mmol/L,pH 7.0)作为T线划膜液体系,羊抗鸡二抗以及呕吐毒素抗原均匀地划于硝酸纤维素膜(NC)上质控线和检测线的位置,C线划膜浓度0.2 mg/mL, T2线划膜浓度0.05 mg/mL, T1线划膜浓度0.15 mg/mL。依次将样品垫、结合垫、NC膜和吸水纸组装切割装入检测卡中。测试结果表明定量值250 μg/kg的抑制率为39.54%。并进一步使用检测卡检测来自国家粮食和物资储备局科学研究院、江苏质检中心以及南京微测生物科技有限公司的18个质控样本,样本类型覆盖玉米、小麦以及小麦制品。与液相色谱法检测出的结果相符,准确度的平均值高达102.41%。该检测方法适合用于现场的快速筛查,具有广阔的市场应用前景。Using the rare element europium as the tracer,a high-sensitivity time-resolved fluorescence rapid quantitative assay card for the detection of deoxynivalenol in grain was developed.DON monoclonal antibody was labeled with EDC of 25μL/mL and antibody of 10μg/mL and sprayed on the binding pad with borate-Borax buffer(12 mmol/L,pH 7.27)system as film diluent.Tris-HCl(10 mmol/L,pH 7.0),as the liquid system of T-line film,was uniformly plotted on the quality control line and detection line of nitrocellulose membrane at the concentration of 0.2 mg/mL on the C line,0.05 mg/mL on the T2 line and 0.15 mg/mL on the T1 line.The sample pad,bonding pad,nitrocellulose membrane and absorbent paper were assembled and cut into the test card in turn.The results showed that the inhibition rate of 250μg/kg was 39.54%.The test card was further used to detect 18 quality control samples from the Academy of Science of the State Food Administration,Jiangsu Quality Inspection Center and Nanjing Micromeasurement Biotechnology Co.,Ltd.The sample types covered corn,wheat and wheat products.The results are consistent with the results obtained by liquid chromatography,and the average accuracy is up to 102.41%.This detection method is suitable for rapid screening in the field and has broad market application prospects.

关 键 词：呕吐毒素 时间分辨荧光微球 快速定量 高灵敏度 

分 类 号：TS207.3[轻工技术与工程—食品科学]