食品与发酵工业 >

2021 , Vol. 47 >Issue 12: 224 - 230

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026976

分析与检测

双模态改良型酶联免疫吸附测定法用于黄曲霉毒素B1的检测研究

  • 张光胤 ,
  • 蔡爱丽 ,
  • 邓放明 ,
  • 赵倩 ,
  • 石星波
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  • 1(食品科学与生物技术湖南省重点实验室,湖南 长沙,410128)
    2(湖南农业大学 食品科学技术学院,湖南 长沙,410128)
硕士(赵倩副教授为通讯作者,E-mail:zhaoqian@hunau.edu.cn)

收稿日期: 2021-02-03

  修回日期: 2021-03-16

  网络出版日期: 2021-07-22

基金资助

国家自然科学基金面上项目(31972155);湖南省自然科学基金(2019JJ40115;2020RC3050);中国博士后基金面上项目(2018M642980);长沙市杰出创新青年基金(kq1905017;kq2009058);国家自然科学基金(32001778);湖南省自然科学基金(2020JJ5249)

收起

An improved bimodal ELISA used for the detection of aflatoxin B1

  • ZHANG Guangyin ,
  • CAI Aili ,
  • DENG Fangming ,
  • ZHAO Qian ,
  • SHI Xingbo
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  • 1(Hunan Provincial Key Laboratory of Food Science and Biotechnology, Changsha 410128, China)
    2(College of Food Science and Technology, Hunan Agricultural University, Changsha 410128, China)

Received date: 2021-02-03

  Revised date: 2021-03-16

  Online published: 2021-07-22

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摘要

食品中真菌毒素的污染问题已经引起了广泛关注,而传统的酶联免疫吸附检测方法(enzyme-linked immunosorbent assay,ELISA)中的灵敏度不能满足痕量检测需求。该文拟对传统ELISA进行改进,建立一种比色与热双模态改良型ELISA用于真菌毒素的检测。使用亲和素化的磁纳米颗粒(magnetic nanoparticles,MNPs)为载体富集辣根过氧化物酶(horseradish peroxidase,HRP),与生物素化的黄曲霉毒素B1核酸适配体(aptamer,Apt)制备复合物(MNPs@Apt@HRP)为检测探针。以核酸适配体与抗体组合作为识别元件,3,3',5,5'-四甲基联苯胺(3,3',5,5'-tetramethylbenzidine,TMB)-H2O2的显色反应和氧化态TMB的光热反应为信号输出,实现了检测黄曲霉毒素B1的高灵敏及高选择性的比色及热信号的双模态检测,检测灵敏度分别达到了2.47×10-12 g/L及3.79×10-13g/L,并将该方法成功应用于部分食品中黄曲霉毒素B1的检测。

关键词： 改良型酶联免疫吸附测定法; 适配体; 磁纳米颗粒; 比色信号; 热信号

本文引用格式

张光胤 , 蔡爱丽 , 邓放明 , 赵倩 , 石星波 . 双模态改良型酶联免疫吸附测定法用于黄曲霉毒素B1的检测研究[J]. 食品与发酵工业, 2021 , 47(12) : 224 -230 . DOI: 10.13995/j.cnki.11-1802/ts.026976

Abstract

The fungal toxins contamination in foodstuffs has aroused broad concern. The moderate sensitivity of traditional enzyme-linked immunosorbent assay (ELISA) cannot meet the requirements of trace detection. This project intends to establish a modified ELISA with colorimetric & thermal dual-modal assay for fungal toxins detection by modification and innovation of traditional ELISA. Avidin magnetic nanoparticles (MNPs) were used as the carrier to enrich horseradish peroxidase (HPR), together with the modification of biotinylated aflatoxin B1(AFB1) aptamers, to prepare a complex (MNPs@Apt@HRP) as detection probe. The combination of nucleic acid aptamer and antibody was used as a recognition element, while taking the color reaction of 3,3',5,5'-tetramethylbenzidine (TMB)-H2O2 and the photothermal reaction of oxidized TMB as signal output to proceed high sensitivity and high selectivity colorimetric & thermal dual-modal detection of AFB1. The detection sensitivity reached 2.47×10-12 g/L and 3.79×10-13 g/L. Thereby, this method could be successfully applied in the detection of AFB1 in partial foodstuffs.

Key words： improved ELISA; aptamer; magnetic nanoparticles; colorimetric signal; thermal signal

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