A Simplified Amplification‐Free Strategy with Lyophilized CRISPR‐CcrRNA System for Drug‐Resistant Salmonella Detection

Author:

Bao Qi1,Sun Jiadi1,Fu Xuran1,Sheng Lina1,Ye Yongli1,Ji Jian1,Zhang Yinzhi1,Wang Jiasheng2,Ping Jianfeng3,Sun Xiulan1ORCID

Affiliation:

1. Laboratory of Food Science and Technology School of Food Science and Technology Collaborative Innovation Center of Food Safety and Quality Control Jiangnan University Wuxi 214122 China

2. Department of Environmental Health Science College of Public Health University of Georgia Athens GA 30602 USA

3. School of Biosystems Engineering and Food Science Zhejiang University Hangzhou 310058 China

Abstract

AbstractDrug resistance in pathogenic bacteria has become a major threat to global health. The misuse of antibiotics has increased the number of resistant bacteria in the absence of rapid, accurate, and cost‐effective diagnostic tools. Here, an amplification‐free CRISPR‐Cas12a time‐resolved fluorescence immunochromatographic assay (AFC‐TRFIA) is used to detect drug‐resistant Salmonella. Multi‐locus targeting in combination crRNA (CcrRNA) is 27‐fold more sensitive than a standalone crRNA system. The lyophilized CRISPR system further simplifies the operation and enables one‐pot detection. Induction of nucleic acid fixation via differentially charged interactions reduced the time and cost required for flowmetric chromatography with enhanced stability. The induction of nucleic acid fixation via differentially charged interactions reduces the time and cost required for flowmetric chromatography with enhanced stability. The platform developed for the detection of drug‐resistant Salmonella has an ultra‐sensitive detection limit of 84 CFU mL−1 within 30 min, with good linearity in the range of 102–106 CFU mL−1. In real‐world applications, spiked recoveries range from 76.22% to 145.91%, with a coefficient of variation less than 10.59%. AFC‐TRFIA offers a cost‐effective, sensitive, and virtually equipment‐independent platform for preventing foodborne illnesses, screening for drug‐resistant Salmonella, and guiding clinical use.

Funder

National Natural Science Foundation of China

Fundamental Research Funds for the Central Universities

Jiangnan University

Publisher

Wiley

Subject

Biomaterials,Biotechnology,General Materials Science,General Chemistry

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