生物技术通报 ›› 2023, Vol. 39 ›› Issue (12): 81-89.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0698

• 技术与方法 • 上一篇    下一篇

基于抗体-适配体夹心生物传感器检测大肠杆菌O157: H7

侯炜辰1(), 叶柯1, 李洁2, 张洋子3, 许文涛3, 朱龙佼3, 李相阳1()   

  1. 1.北京农学院食品科学与工程学院 食品质量与安全北京实验室 农产品有害微生物及农残安全检测与控制北京市重点实验室,北京 102206
    2.中国农业大学食品科学与营养工程学院,北京 100083
    3.中国农业大学营养与健康系 食品精准营养与质量控制教育部重点实验室,北京 100083
  • 收稿日期:2023-07-19 出版日期:2023-12-26 发布日期:2024-01-11
  • 通讯作者: 李相阳,男,副教授,研究方向:食品安全;E-mail: lxy2002cn@163.com
  • 作者简介:侯炜辰,女,硕士研究生,研究方向:食品加工与安全;E-mail: houweichen007@163.com
  • 基金资助:
    国家重点研发计划(2022YFF0607900);北京市景观休闲农业创新团队项目(BAIC09-2023);北京市科技计划(Z221100007122004)

Detection of Escherichia coli O157: H7 Based on Antibody Aptamer Sandwich Biosensor

HOU Wei-chen1(), YE Ke1, LI Jie2, ZHANG Yang-zi3, XU Wen-tao3, ZHU Long-jiao3, LI Xiang-yang1()   

  1. 1. Beijing Laboratory of Food Quality and Safety, Beijing Key Laboratory of Detection and Control of Spoilage Organisms and Pesticide Residue in Agricultural Product, College of Food Science and Engineering, Beijing University of Agriculture, Beijing 102206
    2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083
    3. Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, Beijing 100083
  • Received:2023-07-19 Published:2023-12-26 Online:2024-01-11

摘要:

大肠杆菌O157: H7(Escherichia coli O157:H7)是一种重要的与公共卫生相关的食源性病原体。抗体分子是体液免疫应答中最重要的效应分子,具有多种生物学活性,最主要的生物学功能是与相应抗原特异性结合。适配体是体外合成的较短DNA序列,可通过识别特定区域和靶标特异性结合。利用抗体和适配体的特异性识别功能及免疫磁珠的捕获作用和磁分离,并通过多克隆抗体和裁剪得到的适配体搭建生物传感器,使用实时荧光定量聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction,qPCR)可以实现对靶标在(8×103)-(8×106) CFU/mL范围内的定量检测,检测限为800 CFU/mL。

关键词: 多克隆抗体, 适配体, 裁剪, 免疫磁珠, 大肠杆菌O157: H7

Abstract:

Escherichia coli O157:H7 is an important food borne pathogen related to public health. Antibody molecules are the most important effector molecules in the humoral immune response. They have a variety of biological activities. The main biological function is to specifically bind to the corresponding antigen. Aptamers are shorter DNA sequences synthesized in vitro, which can bind specifically by recognizing specific regions and targets. Based on the specific recognition function of antibody and aptamer, the capture effect and magnetic separation of immunomagnetic beads, and the construction of biosensor by polyclonal antibody and tailoring aptamer, real-time fluorescence quantitative polymerase chain reaction(qPCR)may achieve the detection of target at (8×103)-(8×106) CFU/mL. The limit of detection is 800 CFU/mL for quantitative detection.

Key words: polyclonal antibody, aptamer, tailoring, immunomagnetic beads, Escherichia coli O157: H7