高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (6): 1333.doi: 10.7503/cjcu20130132

• 分析化学 • 上一篇    下一篇

纳米金标记抗体增强SPR检测大肠杆菌O157: H7

刘霞1, 李蓉卓1, 李蕾1, 李文进1, 周春娇2   

  1. 1. 湖南农业大学食品科技学院, 食品科学与生物技术湖南省重点实验室, 长沙 410128;
    2. 湖南农业大学理学院, 长沙 410128
  • 收稿日期:2013-02-04 出版日期:2013-06-10 发布日期:2013-05-15
  • 通讯作者: 刘霞,女,博士,副教授,主要从事食品分析与生物技术的研究.E-mail:liuxiaspr@gmail.com E-mail:liuxiaspr@gmail.com
  • 基金资助:

    国家自然科学基金(批准号: 31201375)和湖南省教育厅优秀青年项目(批准号: 10B050)资助.

Immunoanalysis of E. coli O157: H7 Based on Au Nanoparticles Labelling Antibody Using SPR Biosensor

LIU Xia1, LI Rong-Zhuo1, LI Lei1, LI Wen-Jin1, ZHOU Chun-Jiao2   

  1. 1. Hunan Province Key Laboratory of Food Science and Biotechnology, College of Food Science and Technology, Hunan Agricultural University, Changsha 410128, China;
    2. College of Science, Hunan Agricultural University, Changsha 410128, China
  • Received:2013-02-04 Online:2013-06-10 Published:2013-05-15

摘要:

将金纳米粒子(AuNPs)标记的大肠杆菌O157: H7(E. coli O157: H7)的多克隆抗体(PAb)作为二抗, 采用氨基偶联法将PAb固定在传感器表面作为一抗, 通过三明治方法用双通道表面等离子体子共振(SPR)传感器对E. coli O157: H7 进行检测, 并与SPR直接法检测进行了比较. 结果表明, 直接法的检出限为103 cfu/mL, 线性范围为103~109 cfu/mL; AuNPs增强三明治法的检出限为10 cfu/mL, 线性范围为10~1010 cfu/mL, 灵敏度比直接法提高了100倍, 且具有更宽的检测范围. 本方法不仅检测时间短, 而且具有良好的选择性和重现性.

关键词: 金纳米粒子, 大肠杆菌O157: H7, 表面等离子体子共振, 标记, 检测

Abstract:

The anti-E. coli O157: H7 polyclonal antibodies(PAb) were labelled with Au nanoparticles(AuNPs), which was used as secondary antibodies. The PAb, which was used as primary antibodies, was immobilized on the sensor surface with amino coupling method. The E. coli O157: H7 was detected using direct assay and enhancing sandwich assay based on the two channels surface plasmon resonance(SPR) biosensor. The result indicated a good linear relationship between the SPR signal and logarithmic value of E. coli O157: H7 concentration ranging from 103 cfu/mL to 109 cfu/mL, with the limit of detection(LOD) of 103 cfu/mL for the direct assay. For AuNPs enhancing sandwich assay, the linear range and the limit of detection were determined to be 10-1010 cfu/mL and 10 cfu/mL, respectively. The sensitivity is 100 times higher than that of direct detection. By introducing AuNPs-PAb compound, a wider detection range and lower detection limit could be obtained. This method is rapid, and has good selectivity and reproducibility.

Key words: Au nanoparticle(AuNP), Escherichia coli(E. coli) O157: H7, Surface plasmon resonance(SPR), Label, Detection

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