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A CRISPR/dCas9‐enabled, on‐site, visual, and bimodal biosensing strategy for ultrasensitive and self‐validating detection of foodborne pathogenic bacteria

Yaru LiState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology Tianjin University of Science & Technology Tianjin ChinaJiali QiaoState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology Tianjin University of Science & Technology Tianjin ChinaZhiying ZhaoState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology Tianjin University of Science & Technology Tianjin ChinaQiang ZhangBranch of Tianjin Third Central Hospital Tianjin ChinaWenlu ZhangState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology Tianjin University of Science & Technology Tianjin ChinaShuli ManState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology Tianjin University of Science & Technology Tianjin ChinaShengying YePharmacy Department The 983th Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army Tianjin ChinaKai ChenCollege of Life Science The Innovation Institute of Agricultural Technology Shangrao Normal University Shangrao ChinaLong MaState Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology Tianjin University of Science & Technology Tianjin China

2023en

ABI

Annotatsiya

Abstract It is imperative to develop practicable pathogenic bacteria detection methods. We devised a biosensor for the ultrasensitive detection of Salmonella typhimurium ( S. typhi ), termed as SCOUT‐dCas9 (ultrasensitive, cross‐validating, on‐site, and dUal‐mode test using CRISPR/dCas9). Simply, the species‐specific invA gene of S. typhi was amplified using loop‐mediated isothermal amplification with a biotinylated primer, which can be specifically “pulled down” by “antibody‐like” dCas9‐single guide RNA to form a ternary complex. SYBR Green I and streptavidin‐modified alkaline phosphatase were used to functionalize them to generate fluorescent and colorimetric signals, respectively. With this strategy, the target could be dexterously converted into bimodal signals that were cross‐validated to afford more reliable results. For both modes, SCOUT‐dCas9 was able to detect as low as 1 CFU/mL with a dynamic range from 1 to 10 9 CFU/mL. Lastly, SCOUT‐dCas9 had satisfactory selectivity and was capable of detecting S. typhi ‐contaminated real food samples. SCOUT‐dCas9 provides a robust platform for bacterial detection.

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Identifikatorlar

DOI: 10.1002/fft2.286

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