Collaborative Research: CRISPR-SERS system for rapid and ultrasensitive detection of foodborne bacterial pathogens

细菌病原体及其毒素是食源性疾病最常见的原因。检测复杂基质(如新鲜农产品)中的致病细菌污染物是确保食品安全的重要步骤。检测这些污染物的主要挑战之一是缺乏快速和超灵敏的检测方法。这一多学科项目旨在开发创新的生物传感器，能够快速准确地对大量样本进行致病污染物筛查。该项目与通过Stemette夏令营和工程高中日提供的外联活动相结合，向K-12学生介绍最先进的生物传感器技术。此外，密苏里大学和莱斯大学现有的本科生和研究生课程都纳入了基于该项目成果的新课程单元。本项目包括三个研究目标：(1)设计一个簇状的、有规则间隔的短回文重复序列相关体系(CRISPR-Cas)来检测食源性致病菌；(2)在微流控装置中开发一个超灵敏的SERS体系；(3)将CRISPR和SERS集成在微流控装置中，用于快速、超灵敏和选择性地检测食源性致病菌。这项研究发现、促进和补充了对设计基于CRISPR的下一代生物传感器的基本原则的基本理解。通过将CRISPR与SERS相结合，这项研究还创建了一种新的传感机制，有可能实现具有单拷贝灵敏度和单碱基选择性的检测。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Bacterial pathogens and their toxins are the most common causes of foodborne illnesses. Detecting pathogenic bacterial contaminants in complex matrices (e.g., fresh produce) is an essential step to ensure food safety. One of the major challenges for detecting those contaminants lies in the lack of rapid and ultrasensitive detection methods. This multidisciplinary project aims to develop innovative biosensors that can rapidly and accurately screen a large number of samples for pathogenic contaminants. This project is integrated with outreach activities offered through the STEMette Summer Camp and Engineering High School Day to introduce K-12 students to the state-of-the-art biosensor technology. In addition, new course modules based on the results of this project are incorporated into existing undergraduate and graduate courses at both the University of Missouri and Rice University. This project includes three research objectives: (1) design a clustered, regularly interspaced, short palindromic repeats-associated system (CRISPR-Cas) to detect foodborne bacterial pathogens, (2) develop an ultrasensitive SERS system in a microfluidic device, and (3) integrate CRISPR with SERS in the microfluidic device for rapid, ultrasensitive, and selective detection of foodborne bacterial pathogens. This research discovers, contributes to, and supplements the fundamental understanding of essential principles for engineering CRISPR-based next-generation biosensors. By coupling CRISPR with SERS, this study also creates a new sensing mechanism that has the potential to achieve detection with single-copy sensitivity and single-base selectivity.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.