G-Quadruplex-Probing CRISPR-Cas12 Assay for Label-Free Analysis of Foodborne Pathogens and Their Colonization In Vivo
Autor: | Mohammad Rizwan Khan, Changwei Lei, Kaixiang Zhang, Sha Deng, Xuhan Xia, Qiang He, Boheng Ma, Guiping He, Yun Hu, Ting Zhang, Yunhao Lu, Ruijie Deng |
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Rok vydání: | 2021 |
Předmět: |
Fluid Flow and Transfer Processes
biology business.industry Process Chemistry and Technology Loop-mediated isothermal amplification RNA Bioengineering biology.organism_classification Food safety eye diseases Microbiology Molecular Diagnostic Techniques Salmonella enterica In vivo Animals CRISPR Colonization CRISPR-Cas Systems business Chickens Nucleic Acid Amplification Techniques Instrumentation Gene |
Zdroj: | ACS Sensors. 6:3295-3302 |
ISSN: | 2379-3694 |
Popis: | Foodborne pathogen infection is a key issue of food safety. Herein, we developed a label-free assay for Salmonella enterica (S. enterica) detection based on the G-quadruplex-probing CRISPR-Cas12 system (termed G-CRISPR-Cas), allowing highly sensitive detection of S. enterica and investigation of their colonization in chickens. The introduction of the G-quadruplex probe serving as the substrate of Cas 12a realized a label-free analysis for foodborne pathogens. Due to the amplification process induced by loop-mediated isothermal amplification (LAMP), G-CRISPR-Cas assay can detect S. enterica as low as 20 CFU. Specificity for pathogenic gene detection was guaranteed by the dual recognition process via LAMP primers and Cas 12a-guided RNA binding. The G-CRISPR-Cas assay was applied to explore S. enterica colonization in the intestinal tract and organs of chickens and showed the risk of S. enterica infection outside of the intestinal tract. The G-CRISPR-Cas assay is promising for on-site diagnosis of the infection or contamination of foodborne pathogens outside the laboratories, such as abattoirs and markets. |
Databáze: | OpenAIRE |
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