化学学报 ›› 2012, Vol. 70 ›› Issue (11): 1304-1308.DOI: 10.6023/A1201033 上一篇    下一篇

研究论文

纳米金增强SPR 检测产毒赭曲霉中PKS 基因的特异性碱基

李迎, 林钊, 李蓉卓, 刘霞   

  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
  • 收稿日期:2012-01-03 修回日期:2012-03-06 出版日期:2012-06-14 发布日期:2012-03-09
  • 通讯作者: 刘霞 E-mail:liuxiaspr@gmail.com
  • 基金资助:

    湖南省教育厅优秀青年项目(No. 10B050)、长沙市科技计划重点项目(No. K1005181-21)和湖南省研究生创新基金(No. CX2010B281)资助项目.

Detection for Polyketide Synthase Gene of Aspergillus ochraceus based on Au Nanoparticles Enhancing SPR Biosensor

Li Ying, Lin Zhao, Li Rongzhuo, Liu Xia   

  1. College of Food Science and Technology, Hunan Agricultural University, Hunan Province Key Laboratory of Food Science and Biotechnology, Changsha 410128
  • Received:2012-01-03 Revised:2012-03-06 Online:2012-06-14 Published:2012-03-09
  • Supported by:

    Project supported by the Scientific Research Fund of Hunan Provincial Education Department (No. 10B050), Key Project of Science and Technology Plan of Changsha (No. K1005181-21) and Graduate Innovation Fund of Hunan Province (No. CX2010B281).

基于双通道表面等离子体子共振(surface plasmon resonance, SPR)传感器, 分别采用直接法和金纳米粒子作为传感层的方法, 通过检测赭曲霉毒素A (ochratoxin A, OTA)合成初期阶段表达的聚酮合成酶(polyketide synthase, PKS)基因的25 个特异性碱基的寡核苷酸链, 建立了一种高灵敏、间接检测OTA 的新方法. 同时考察了6-巯基己醇(6-mercapto-1-hexanol, MCH)作为封闭液对SPR 响应信号的影响. 结果表明, 直接法的检测限为12.5 nmol/L, MCH 的加入可使响应信号有所增强, 使用金纳米粒子作为传感层的检测下限为0.25 nmol/L, 与直接法相比较灵敏度提高了50倍, 与以往使用金纳米粒子标记抗体或抗原相比, 其作为传感层也能大大提高SPR 检测灵敏度, 且操作简单易行.

关键词: 表面等离子体子共振, 金纳米粒子, 赭曲霉毒素A, 聚酮合成酶基因

The surface plasmon resonance (SPR) method is established for indirect detection of the ochratoxin A. The 25-base oligonucleotide sequence of polyketide synthase (PKS) gene which is expressed during early stage of ochratoxin A synthesis, was detected respectively using direct assay and Au nanoparticles as sensing membrane assay based on the two channels surface plasmon resonance (SPR) biosensor. At the same time, influence of 6-mercapto-1-hexanol (MCH) blocking on SPR signal was also studied. As a result, the limit of detection (LOD) of the direct assay was 12.5 nmol/L. And the SPR signal was enhanced with 6-mercapto-1-hexanol (MCH) blocking. With Au nanoparticles as a sensing membrane, LOD was 0.25 nmol/L, sensitivity was 50 times higher than that of direct detection. SPR signal could be dramatically enhanced with Au nanoparticles as sensing membrane instead of as label, and the operation was simple.

Key words: surface plasmon resonance (SPR), Au nanoparticles, ochratoxin A, polyketide synthase (PKS)