CRISPR-based approaches for the detection of antimicrobial-resistant genes

The food industry has been seeking more advanced detection methods, and the emerging technology of biosensing based on clustered regularly interspaced short palindromic repeats (CRISPR) may hold the key to addressing this need. Here, we evaluate the current state of CRISPR technology in the realm of food safety, including the various applications and targets of each Cas effector. Although still in its early stages, CRISPR/Cas-based sensors have shown great promise and could become the new standard in food safety. Additionally, this technology has potential for use in other areas of the food industry, such as detecting antimicrobial resistant (AMR) bacteria. To this end, we developed a novel PCR-coupled CRISPR/Cas12a-based fluorescence assay capable of detecting bacteria that produce New Delhi metallo-β-lactamase (NDM). The CRISPR/Cas12a system, thanks to its specifically designed gRNA, demonstrated remarkable specificity at the single-base pair level, specifically targeting the NDM-producing genes in bacteria. Our assay demonstrated superior performance compared to previously developed real-time PCR assays and addressed important weaknesses of conventional amplification-based detection methods, such as false positives and SNP discrimination. With further research, CRISPR/Cas-based sensors have the potential to become the new gold standard for detecting AMR genes, ultimately contributing to mitigating the global threat of AMR.

Name thesis:degree_name

M.S.

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Food Science & Human Nutrition

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2023

Author dc:creator

• Shin, Jiyong

Contributors dc:contributor

• Wang, Yi-Cheng
• Jin, Yong-Su
• Miller, Michael

• Fluorescent Assay
• Crispr Biosensor
• Cas12a
• Nucleic Acid Detection
• Food Safety
• Multi-drug-resistant

Statement dc:rights

• Copyright 2023 by Jiyong Shin

Language dc:language

en, eng

Handle dc:identifier

https://hdl.handle.net/2142/122188

OAI record oai:identifier

https://www.ideals.illinois.edu/oai-pmh?verb=GetRecord&metadataPrefix=etdms&identifier=oai%3Awww.ideals.illinois.edu%3A2142%2F122188