SPR生物传感器高通量检测多种食源性 致病菌的研究

High-throughput detection for multiplexed food-borne pathogen using surface plasmon resonance biosensor

中文关键词:  表面等离子共振技术  生物芯片  高通量  食源性致病菌

英文关键词:surface plasmon resonance technology  biochip  high throughput  foodborne pathogens

中文摘要:

      目的 建立一种基于表面等离子共振技术 (Surface Plasmon Resonance technology, SPR)原理的生物传感器方法, 实现11种常见致病菌的高通量检测。方法 用表面自组装技术(self-assembled monolayer, SAM)在金膜表面引入羧基, 然后通过氨-羧基反应将5’端标记有氨基的探针固定在芯片表面, 经多重PCR扩增出的产物变性、杂交、生物素-亲和素系统信号放大等步骤完成目的片段的检测。对所建立的多重PCR-SPR检测方法的灵敏性、特异性等技术指标进行评价。结果 本研究建立的多重PCR-SPR检测方法具有良好的灵敏性, 该芯片检测11种常见致病菌的灵敏性均达到103~104 cfu/mL, 能满足现有检测要求。通过对实验室保存的14株标准菌株和沙门氏菌等90份样品分离株进行检测, 结果显示该检测方法具有较好的特异性, 无交叉反应。结论 本研究建立的多重PCR-SPR 检测系统具有良好的灵敏性和特异性, 完全适用于11种致病菌的高通量检测。该检测方法检测时间短、检测成本低、减少了人为因素的影响, 具有良好的应用前景。

英文摘要:

      Objective A biosensor based on surface plasmon resonance (SPR) principle was developed for food-borne pathogen bacteria detection, the method achieved a high-throughput detection for 11 kinds of bacteria. Methods The chip surface was modified by self-assembled monolayer (SAM), and carboxyl group was introduced. The reaction of the carboxyl and amino leading to the probes were arrayed on the chips, and the biotin-avidin system was introduced to enhance the signal after hybridization. We evaluated the sensitivity, the specificity and other technical indicators by multiple PCR-SPR method. Results The multiplex PCR-SPR method showed a good sensitivity. The sensitivity of 11 kinds pathogenic bacteria reached 103~104 cfu/mL, which can meet the requirements of existing detection. Detection of 14 Standard strains and Salmonella strains were isolated from 90 samples. The results showed that the method had a good specificity without cross reaction. Conclusion In this study, a detection method for 11 kinds of common foodborne pathogens was established by multiplex PCR-SPR system. It has a short detection time, a low detection cost, a reduction of the influence of manual factors, and will have a good application prospect.