2026-09-23T10:36:19Z https://www.ideals.illinois.edu/oai-pmh

oai:www.ideals.illinois.edu:2142/122188 2024-03-02 col_2142_5131 col_2142_14781 com_2142_5130 com_2142_687 com_2142_397

Wang, Yi-Cheng Jin, Yong-Su Miller, Michael 2023-05 application/pdf en text Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2025-05-01 The student, Jiyong Shin, accepted the attached license on 2023-04-27 at 14:28. The student, Jiyong Shin, submitted this Thesis for approval on 2023-04-27 at 14:38. This Thesis was approved for publication on 2023-05-01 at 09:30. DSpace SAF Submission Ingestion Package generated from Vireo submission #19195 on 2024-03-01 at 13:47:33 Fluorescent Assay CRISPR-based approaches for the detection of antimicrobial-resistant genes Shin, Jiyong 2023-05-01 Crispr Biosensor Cas12a Nucleic Acid Detection Food Safety Multi-drug-resistant 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. Text eng https://hdl.handle.net/2142/122188 Copyright 2023 by Jiyong Shin M.S. Thesis Food Science & Human Nutrition University of Illinois at Urbana-Champaign Food Science & Human Nutrition