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Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform

Yersinia enterocolitica is a dangerous foodborne human pathogen that mainly causes gastroenteritis. Ideal methods for the detection of pathogens in food should be rapid, sensitive, specific, and cost effective. To this end, novel in vitro nucleic acid identification methods based on clustered, regul...

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Autores principales: Xiao, Yiran, Ren, Honglin, Hu, Pan, Wang, Yang, Wang, Han, Li, Yansong, Feng, Kai, Wang, Cong, Cao, Qi, Guo, Yuxi, Liu, Zengshan, Lu, Shiying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9318358/
https://www.ncbi.nlm.nih.gov/pubmed/35885403
http://dx.doi.org/10.3390/foods11142160
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author Xiao, Yiran
Ren, Honglin
Hu, Pan
Wang, Yang
Wang, Han
Li, Yansong
Feng, Kai
Wang, Cong
Cao, Qi
Guo, Yuxi
Liu, Zengshan
Lu, Shiying
author_facet Xiao, Yiran
Ren, Honglin
Hu, Pan
Wang, Yang
Wang, Han
Li, Yansong
Feng, Kai
Wang, Cong
Cao, Qi
Guo, Yuxi
Liu, Zengshan
Lu, Shiying
author_sort Xiao, Yiran
collection PubMed
description Yersinia enterocolitica is a dangerous foodborne human pathogen that mainly causes gastroenteritis. Ideal methods for the detection of pathogens in food should be rapid, sensitive, specific, and cost effective. To this end, novel in vitro nucleic acid identification methods based on clustered, regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) endonuclease have received increasing attention. In this study, a simple, visual, and ultrasensitive method, based on CRISPR/Cas12a with recombinase polymerase amplification (RPA), was developed for the detection of Y. enterocolitica. The results show that a specific attachment invasion locus gene (ail) can be rapidly detected using a CRISPR/Cas12a-RPA-based system. Application of the method to raw pork, which was artificially infected with Y. enterocolitica, achieved an estimated detection limit of 1.7 CFU/mL in less than 45 min, and this was 100 times lower compared with qPCR. The results indicated that the CRISPR/Cas12a-RPA system has good potential for monitoring pathogenic Y. enterocolitica in the chilled meat supply chain.
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spelling pubmed-93183582022-07-27 Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform Xiao, Yiran Ren, Honglin Hu, Pan Wang, Yang Wang, Han Li, Yansong Feng, Kai Wang, Cong Cao, Qi Guo, Yuxi Liu, Zengshan Lu, Shiying Foods Article Yersinia enterocolitica is a dangerous foodborne human pathogen that mainly causes gastroenteritis. Ideal methods for the detection of pathogens in food should be rapid, sensitive, specific, and cost effective. To this end, novel in vitro nucleic acid identification methods based on clustered, regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) endonuclease have received increasing attention. In this study, a simple, visual, and ultrasensitive method, based on CRISPR/Cas12a with recombinase polymerase amplification (RPA), was developed for the detection of Y. enterocolitica. The results show that a specific attachment invasion locus gene (ail) can be rapidly detected using a CRISPR/Cas12a-RPA-based system. Application of the method to raw pork, which was artificially infected with Y. enterocolitica, achieved an estimated detection limit of 1.7 CFU/mL in less than 45 min, and this was 100 times lower compared with qPCR. The results indicated that the CRISPR/Cas12a-RPA system has good potential for monitoring pathogenic Y. enterocolitica in the chilled meat supply chain. MDPI 2022-07-21 /pmc/articles/PMC9318358/ /pubmed/35885403 http://dx.doi.org/10.3390/foods11142160 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Xiao, Yiran
Ren, Honglin
Hu, Pan
Wang, Yang
Wang, Han
Li, Yansong
Feng, Kai
Wang, Cong
Cao, Qi
Guo, Yuxi
Liu, Zengshan
Lu, Shiying
Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform
title Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform
title_full Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform
title_fullStr Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform
title_full_unstemmed Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform
title_short Ultra-Sensitive and Rapid Detection of Pathogenic Yersinia enterocolitica Based on the CRISPR/Cas12a Nucleic Acid Identification Platform
title_sort ultra-sensitive and rapid detection of pathogenic yersinia enterocolitica based on the crispr/cas12a nucleic acid identification platform
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9318358/
https://www.ncbi.nlm.nih.gov/pubmed/35885403
http://dx.doi.org/10.3390/foods11142160
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