Escherichia coli O157:H7的表面增强拉曼光谱检测方法建立
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陕西科技大学食品科学与工程学院,陕西 西安,710021

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王盼雪(1988—),女,博士,副教授,硕士研究生导师,主要从事食品质量与安全检测研究。E-mail:wangpanxue@sust.edu.cn

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国家自然科学基金青年基金项目(32102063);陕西科技大学高水平博士人才科研启动项目(2017BJ-51)。


Establishment of Surface-enhanced Raman Spectroscopy for Detection of Escherichia coli O157:H7
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School of Food Science and Engineering, Shaanxi University of Science and Technology, Xi’an 710021, China

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    摘要:

    【目的】提高大肠杆菌(Escherichia coliE. coli) O157:H7检测的特异性和灵敏度。【方法】基于磁分离和信号放大策略,结合表面增强拉曼光谱(SERS)技术建立了一种特异性好、灵敏度高的E. coli O157:H7检测方法。以适配体为识别元件,联合磁珠和聚苯乙烯(PS)微球,分别构建分离探针MB@Apt和信号放大型拉曼标签4-MBN@PS@Apt。【结果】当E. coli O157:H7存在时,MB@Apt、4-MBN@PS@Apt与E. coli O157:H7可形成MB@Apt/E. coli O157:H7/4-MBN@PS@Apt“三明治”结构复合物,该复合物可借助外源磁场实现快速分离和富集;加入四氢呋喃(THF)后,拉曼标签上负载的低生物背景拉曼报告物4-MBN可被快速释放。4-MBN在1 073.9 cm-1处的相对拉曼强度与E. coli O157:H7浓度的对数值在1×103~1×108 CFU/mL的线性关系良好,决定系数(R2)为0.98。该方法对E. coli O157:H7的检测限(LOD)为312 CFU/mL,在自来水和牛奶样本中的检测回收率为61.87%~135.55%,相对标准偏差(RSD)为0.27%~5.28%。【结论】该方法对E. coli O157:H7的检测灵敏度较高,为食源性致病菌的快速检测提供了新思路。

    Abstract:

    [Objective] This study aims to improve the detection specificity and sensitivity for Escherichia coli (E. coli) O157:H7.[Method] A highly specific and sensitive method for the detection of E. coli O157:H7 was established based on magnetic separation and signal amplification strategies combined with surface-enhanced Raman spectroscopy (SERS). Aptamers were used as recognition elements and combined with magnetic beads and polystyrene (PS) microspheres to construct a separation probe (MB@Apt) and a signal amplification Raman tag (4-MBN@PS@Apt), respectively.[Result] In the presence of E. coli O157:H7, MB@Apt and 4-MBN@PS@Apt formed a “sandwich” structured composite (MB@Apt/E. coli O157:H7/4-MBN@PS@Apt), which could be rapidly separated and enriched by an external magnet. After tetrahydrofuran (THF) was added, the low biological background Raman reporter 4-MBN in the Raman tag was quickly released. The relative Raman intensity of 4-MBN at 1 073.9 cm-1 showed a good linear relationship with the logarithm of E. coli O157:H7 concentration in the range of 1×103 to 1×108 CFU/mL, with a determination coefficient (R2) of 0.98. The established method showed a limit of detection (LOD) of 312 CFU/mL for E. coli O157:H7, the recovery rates ranging from 61.87% to 135.55% in tap water and milk samples, and relative standard deviations (RSD) between 0.27% and 5.28%.[Conclusion] This method exhibited high detection sensitivity for E. coli O157:H7 and provided a new approach for rapid detection of foodborne pathogenic bacteria.

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王盼雪,李岑,李香,刘莹,李国梁.Escherichia coli O157:H7的表面增强拉曼光谱检测方法建立[J].食品与生物技术学报,2025,(6):97-105.

WANG Panxue, LI Cen, LI Xiang, LIU Ying, LI Guoliang. Establishment of Surface-enhanced Raman Spectroscopy for Detection of Escherichia coli O157:H7[J]. Journal of Food Science and Biotechnology,2025,(6):97-105.

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  • 收稿日期:2023-07-05
  • 最后修改日期:2023-09-26
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  • 在线发布日期: 2025-10-11
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