Monitoring Pathogenic Viable E. coli O157:H7 in Food Matrices Based on the Detection of RNA Using Isothermal Amplification and a Paper-Based Platform

In recent years, the number of product recalls and contamination incidents involving pathogenic bacteria has significantly increased, and the ensuing infections continue to be an ongoing problem for public health and agriculture. Due to the widespread impact of these pathogens, there is a critical n...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2022-02, Vol.94 (5), p.2485-2492
Main Authors: Petrucci, Sabrina, Costa, Connor, Broyles, David, Kaur, Avinash, Dikici, Emre, Daunert, Sylvia, Deo, Sapna K
Format: Article
Language:English
Subjects:
Analytical chemistry
Animals
Bacteria
Cattle
Cell culture
Chemistry
E coli
Escherichia coli
Escherichia coli O157 - genetics
Food contamination
Food Contamination - analysis
Food Microbiology
Monitoring
Pathogens
Product recalls
Public health
Recombinase
Reverse transcription
RNA
Sample preparation
Spinacia oleracea
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Summary:In recent years, the number of product recalls and contamination incidents involving pathogenic bacteria has significantly increased, and the ensuing infections continue to be an ongoing problem for public health and agriculture. Due to the widespread impact of these pathogens, there is a critical need for rapid, on-site assays that can provide rapid results. In this work, we demonstrate the development of a rapid and simple test based on the combination of reverse transcription with recombinase polymerase amplification followed by lateral flow strip detection of viable Escherichia coli O157:H7 cells by detecting the RNA of the pathogen. The optimized method can be performed for approximately 2 h with a detection limit of 10 CFU/mL of E. coli O157:H7 in buffer, spinach, and ground beef samples. Our assay is sensitive, detecting only E. coli O157:H7 and not nonpathogenic E. coli or other similar pathogens. This strategy was able to distinguish viable from nonviable bacteria and more significantly was able to detect viable but nonculturable bacteria, which is a major issue when using culture-based methods for monitoring pathogenic bacteria. An important advantage of this test is that it can provide timely identification and removal of contaminated consumables prior to distribution without an extensive sample preparation.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.1c04305