食品科学 ›› 2021, Vol. 42 ›› Issue (24): 311-317.doi: 10.7506/spkx1002-6630-20201217-211

• 安全检测 • 上一篇    

基于金纳米颗粒信号放大ELISA检测食品中恩诺沙星

李坚,张富源,刘敏轩,刘若冰,王向红   

  1. (河北农业大学食品科技学院,河北 保定 071000)
  • 发布日期:2021-12-30
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2016YFD0401101)

Development and Application of a Gold Nanoparticle-Based Signal Amplification Enzyme Linked Immunosorbent Assay for the Detection of Enrofloxacin in Food Samples

LI Jian, ZHANG Fuyuan, LIU Minxuan, LIU Ruobing, WANG Xianghong   

  1. (College of Food Science and Technology, Hebei Agricultural University, Baoding 071000, China)
  • Published:2021-12-30

摘要: 以金纳米颗粒(gold nanoparticles,AuNPs)为信号放大载体,利用辣根过氧化物酶(horseradish peroxidase,HRP)标记的二抗为信号探针,建立一种基于AuNPs的信号放大酶联免疫检测方法(AuNPs signal amplification enzyme-linked immunosorbent assay,AuNPs-HRP-IgG ic-ELISA),检测食品中恩诺沙星(enrofloxacin,ENR)。该方法对ENR的检测限(IC15)为5×10-4?ng/mL,半数抑制浓度(IC50)为0.24?ng/mL,检测范围为0.16~500?ng/mL,与建立的传统ELISA方法(IC50=8.76?ng/mL)相比,显著提高了检测灵敏度,并且该方法与环丙沙星、氧氟沙星、诺氟沙星交叉反应率均小于0.1%,具有良好的特异性。该AuNPs-HRP-IgG?ic-ELISA方法的实用性得到了样品添加回收实验和商业化ELISA检测试剂盒的验证,其在牛奶样品中的加标回收率可达80.52%~102.66%,适用于实际牛奶样本中ENR的快速灵敏检测,也为建立其他食品危害物质的精准检测技术开发提供参考。

关键词: 恩诺沙星;金纳米颗粒;辣根过氧化物酶;酶联免疫检测方法;灵敏度

Abstract: In this study, a signal amplification enzyme-linked immunosorbent assay (AuNPs-HRP-IgG ic-ELISA) to detect enrofloxacin (ENR) in foods was established using gold nanoparticles (AuNPs) as a signal amplification carrier and horseradish peroxidase (HRP)-labeled secondary antibody as a signal probe. The detection limit (IC15) of this method for ENR was 5 × 10-4 ng/mL, the half maximal inhibitory concentration (IC50) was 0.24 ng/mL, and the detection range was 0.16–500 ng/mL. Compared with the traditional ELISA method (IC50 = 8.76 ng/mL), the sensitivity was significantly improved. The cross-reactivity of this method was less than 0.1% toward ciprofloxacin, ofloxacin, and norfloxacin, indicating good specificity. The feasibility of the AuNPs-HRP-IgG ic-ELISA was validated in terms of spiked recoveries in comparison with a commercial ELISA kit. The recoveries for spiked milk samples was 80.52%–102.66%, suggesting that this method could be suitable for the rapid and sensitive detection of ENR in actual milk samples. This study may also provide a new way to develop detection techniques for other related hazardous substances in foods.

Key words: enrofloxacin; gold nanoparticles; horseradish peroxidase; enzyme linked immunosorbent assay; sensitivity

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