利用时间分辨荧光免疫层析法同时定量检测玉米中的伏马毒素B1、B2和B3
Application of Time-resolved Fluorescence Immunoassay to Concurrently Quantify Fumonisin B1,B2 and B3 in Corn
张佳林 1马立才 2侯璐 3孟和那仁 4温凯 5侯晓林3
作者信息
- 1. 北京农学院动物科学技术学院,北京 102206;北京维德维康生物技术有限公司,北京 100095
- 2. 北京维德维康生物技术有限公司,北京 100095
- 3. 北京农学院动物科学技术学院,北京 102206
- 4. 赤峰市农牧业综合行政执法支队,内蒙古赤峰 024000
- 5. 中国农业大学动物医学院,北京 100193
- 折叠
摘要
建立了一种可同时定量检测玉米中伏马毒素B1、B2和B3的时间分辨荧光免疫层析方法.利用活化酯法制备时间分辨荧光微球和伏马毒素抗体的偶联物,将伏马毒素抗原包被到硝酸纤维素膜上作为T线,通过优化不同T线包被条数,最终建立伏马毒素竞争性免疫层析方法.并对所建立方法的灵敏度、特异性、准确度、精密度和与国标方法分析结果的相关性进行评价.包被两条T线建立的检测方法检出限为107.68～168.28 ng/kg,定量限为283.46～444.63 μg/kg;与伏马毒素B1、伏马毒素B2和伏马毒素B3的交叉反应率分别为100％、85.59％和72.72％,与其它5种常见的真菌毒素均无明显交叉;加标回收率范围88.37％～117.42％,变异系数小于10％;该方法与国标方法GB 5009.240-2016中规定的免疫亲和柱净化-高效液相色谱法(IAC-HPLC)检测结果的符合度在92.17％～107.21％之间.建立的时间分辨荧光免疫层析检测方法能满足对玉米中伏马毒素B1、B2和B3的现场快速定量检测.
Abstract
This study aimed to establish a time-resolved fluorescence immunoassay method for the simultaneous quantitative detection of fumonisin B1,B2 and B3 in corn.The EDC/NHS active ester method was used to prepare time-resolved fluorescent microspheres and fumonisin antibody conjugates.Fumonisin antigen was immobilized on a nitrocellulose membrane as the test line.By optimizing the number of test line,a competitive immunoassay method for detecting fumonisin was eventually established.Subsequently,the sensitivity,specificity,accuracy,precision,and correlation with the national standard method were evaluated for the established method.The detection limits of the established method by using two test line pattern were 107.68～168.28 pg/kg,and the quantification limits were 283.46～444.63 pg/kg.The cross-reactivity with fumonisin B1,fumonisin B2 and fumonisin B3 were 100％,85.59％and 72.72％,respectively,and no significant cross-reactivity was observed with five other common mycotoxins.The recovery rates ranged from 88.37％to 117.42％,and the coefficients of variation were lower than 10％.The conformity of the results obtained by the established method,as compared to the immunoaffinity column purification-high performance liquid chromatography(IAC-HPLC)method specified in the national standard GB 5009.240-2016,fell within the range of 92.17％to 107.21％.The established time-resolved fluorescence immunoassay method meets the requirements for rapid on-site quantitative detection of fumonisin B1,B2 and B3 in corn.
关键词
时间分辨荧光微球/伏马毒素B1/伏马毒素B2/伏马毒素B3/玉米/定量检测Key words
time-resolved fluorescent microsphere/fumonisin B1/fumonisin B2/fumonisin B3/corn/quantitative detection引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据