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Establishment of a Simple, Sensitive, and Specific Detection Method Based on Recombinase-Aided Amplification Combined with dsDNA-Specific Nucleases.

作者信息

Zhou Changyu, Zhao Yu, Guo Boyan, Yang Ming, Xu Qiang, Lei Changwei, Wang Hongning

机构信息

Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610017, China.

Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Chengdu 610064, China.

出版信息

Foods. 2024 Apr 29;13(9):1380. doi: 10.3390/foods13091380.


DOI:10.3390/foods13091380
PMID:38731750
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11083397/
Abstract

is a common foodborne pathogen that can cause food poisoning, posing a serious threat to human health. Therefore, quickly, sensitively, and accurately detecting is crucial to ensuring food safety. For the gene, we designed Recombinase-aided amplification (RAA) primers and dsDNA-specific nuclease (DNase) probes. The ideal primer and probe combination was found when conditions were optimized. Under UV light, a visual detection technique (RAA-dsDNase) was developed. Additionally, the RAA-dsDNase was modified to further reduce pollution hazards and simplify operations. One-pot RAA-dsDNase-UV or one-pot RAA-dsDNase-LFD was developed as a detection method, using UV or a lateral flow dipstick (LFD) for result observation. Among them, one-pot RAA-dsDNase and one-pot RAA-dsDNase-LFD had detection times of 50 min and 60 min, respectively, for detecting genomic DNA. One-pot RAA-dsDNase-UV had a detection limit of 10 copies/μL and 10 CFU/mL, while one-pot RAA-dsDNase-LFD had a sensitivity of 10 copies/μL and 10 CFU/mL. One-pot RAA-dsDNase-UV and one-pot RAA-dsDNase-LFD assays may identify 17 specific serovars witho ut causing a cross-reaction with the remaining 8 bacteria, which include . Furthermore, in tissue and milk samples has been reliably detected using both approaches. Overall, the detection method developed in this study can quickly, sensitively, and accurately detect , and it is expected to become an important detection tool for the prevention and control of in the future.

摘要
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/d950999f85d2/foods-13-01380-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/c96d11051ff4/foods-13-01380-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/709f3cdb0a07/foods-13-01380-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/9ef9a2ca9b38/foods-13-01380-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/60a9b1a2d96e/foods-13-01380-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/6f563f3f6655/foods-13-01380-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/d950999f85d2/foods-13-01380-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/c96d11051ff4/foods-13-01380-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/709f3cdb0a07/foods-13-01380-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/9ef9a2ca9b38/foods-13-01380-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/60a9b1a2d96e/foods-13-01380-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/6f563f3f6655/foods-13-01380-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/11083397/d950999f85d2/foods-13-01380-g006.jpg

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Establishment of a Simple, Sensitive, and Specific Detection Method Based on Recombinase-Aided Amplification Combined with dsDNA-Specific Nucleases.

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引用本文的文献

[1]
Molecular methods for rapid detection and identification of foodborne pathogenic bacteria.

World J Microbiol Biotechnol. 2025-5-15

[2]
A rapid and visual detection assay for Senecavirus A based on recombinase-aided amplification and lateral flow dipstick.

Front Cell Infect Microbiol. 2024

本文引用的文献

[1]
Corrigendum to "Establishment of an ultrasensitive and visual detection platform for Neospora caninum based-on the RPA-CRISPR/Cas12a system" [Talanta vol. (2024) 269/125413].

Talanta. 2024-5-1

[2]
CeO nanozyme mediated RPA/CRISPR-Cas12a dual-mode biosensor for detection of invA gene in Salmonella.

Biosens Bioelectron. 2024-3-1

[3]
An electrochemical biosensor based on phage-encoded protein RBP 41 for rapid and sensitive detection of Salmonella.

Talanta. 2024-4-1

[4]
A gas-driven capillary based on the synergy of the catalytic and photothermal effect of PB@Au for Salmonella typhimurium detection.

Talanta. 2024-3-1

[5]
A multiplex real-time RT-PCR system to simultaneously diagnose 16 pathogens associated with swine respiratory disease.

J Appl Microbiol. 2023-11-1

[6]
A new method for the detection of Mycobacterium tuberculosis based on the CRISPR/Cas system.

BMC Infect Dis. 2023-10-11

[7]
Advances in the application of recombinase-aided amplification combined with CRISPR-Cas technology in quick detection of pathogenic microbes.

Front Bioeng Biotechnol. 2023-8-31

[8]
Salmonella detection with LAMP and qPCR and identification of serovars of interest by multiplex qPCR in poultry carcasses.

Braz J Microbiol. 2023-9

[9]
Phage amplification coupled with loop-mediated isothermal amplification (PA-LAMP) for same-day detection of viable Salmonella Enteritidis in raw poultry meat.

Food Microbiol. 2023-10

[10]
Ultrasensitive and Rapid Visual Detection of O157:H7 Based on RAA-CRISPR/Cas12a System.

Biosensors (Basel). 2023-6-16

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