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。检测复杂基质中的致病性细菌污染物（例如，新鲜农产品）是确保食物安全的重要步骤。检测这些污染物的主要挑战之一在于缺乏快速和超灵敏的检测方法。这个多学科项目旨在开发创新的生物传感器，可以快速，准确地筛选大量的致病性污染物样品。 该项目与通过STEMette夏令营和工程高中日提供的外展活动相结合，向K-12学生介绍最先进的生物传感器技术。此外，根据这一项目的成果，新的课程单元已纳入密苏里州大学和莱斯大学现有的本科生和研究生课程。该项目包括三个研究目标：（1）设计一个聚类的，规则间隔的，短回文重复序列相关系统（CRISPR-Cas）来检测食源性细菌病原体，（2）在微流控装置中开发一个超灵敏的Sers系统，（3）将CRISPR与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细菌病原体。这项研究发现，有助于并补充了对基于CRISPR的下一代生物传感器工程基本原理的基本理解。通过将CRISPR与Sers相结合，该研究还创造了一种新的传感机制，有可能实现单拷贝灵敏度和单碱基选择性的检测。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值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.