Cargando…

A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food

Shigella and Bacillus cereus are two common foodborne pathogens that cause intestinal diseases and seriously affect human life and health. Traditional microbiological culture methods are time-consuming and laborious, and polymerase chain reaction (PCR)-based methods rely on expensive thermal cyclers...

Descripción completa

Detalles Bibliográficos
Autores principales: Xiang, Shuna, Zhang, Hanyue, Cha, Xiaoyan, Lin, Yingting, Shang, Ying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10178236/
https://www.ncbi.nlm.nih.gov/pubmed/37174427
http://dx.doi.org/10.3390/foods12091889
_version_ 1785040813562003456
author Xiang, Shuna
Zhang, Hanyue
Cha, Xiaoyan
Lin, Yingting
Shang, Ying
author_facet Xiang, Shuna
Zhang, Hanyue
Cha, Xiaoyan
Lin, Yingting
Shang, Ying
author_sort Xiang, Shuna
collection PubMed
description Shigella and Bacillus cereus are two common foodborne pathogens that cause intestinal diseases and seriously affect human life and health. Traditional microbiological culture methods are time-consuming and laborious, and polymerase chain reaction (PCR)-based methods rely on expensive thermal cyclers and lengthy reaction times. In this study, on the basis of the specific gene ipaH7 of Shigella and the virulence gene nheABC of B. cereus, a duplex detection system was established for the first time by using the recombinase polymerase amplification technique (D-RPA). After optimization, D-RPA could be effectively amplified at 42 °C for 25 min with excellent specificity, and the detection limits of D-RPA for Shigella and B. cereus in artificially contaminated samples were 2.7 × 10(1) and 5.2 × 10(2) CFU/mL, respectively. This study provides a certain research basis for multiple detection with RPA, an isothermal amplification technology. Furthermore, it lays a good foundation for high-throughput rapid detection of foodborne pathogens.
format Online
Article
Text
id pubmed-10178236
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-101782362023-05-13 A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food Xiang, Shuna Zhang, Hanyue Cha, Xiaoyan Lin, Yingting Shang, Ying Foods Article Shigella and Bacillus cereus are two common foodborne pathogens that cause intestinal diseases and seriously affect human life and health. Traditional microbiological culture methods are time-consuming and laborious, and polymerase chain reaction (PCR)-based methods rely on expensive thermal cyclers and lengthy reaction times. In this study, on the basis of the specific gene ipaH7 of Shigella and the virulence gene nheABC of B. cereus, a duplex detection system was established for the first time by using the recombinase polymerase amplification technique (D-RPA). After optimization, D-RPA could be effectively amplified at 42 °C for 25 min with excellent specificity, and the detection limits of D-RPA for Shigella and B. cereus in artificially contaminated samples were 2.7 × 10(1) and 5.2 × 10(2) CFU/mL, respectively. This study provides a certain research basis for multiple detection with RPA, an isothermal amplification technology. Furthermore, it lays a good foundation for high-throughput rapid detection of foodborne pathogens. MDPI 2023-05-04 /pmc/articles/PMC10178236/ /pubmed/37174427 http://dx.doi.org/10.3390/foods12091889 Text en © 2023 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
Xiang, Shuna
Zhang, Hanyue
Cha, Xiaoyan
Lin, Yingting
Shang, Ying
A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food
title A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food
title_full A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food
title_fullStr A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food
title_full_unstemmed A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food
title_short A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of Shigella and Bacillus cereus in Food
title_sort new duplex recombinase polymerase amplification (d-rpa) method for the simultaneous and rapid detection of shigella and bacillus cereus in food
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10178236/
https://www.ncbi.nlm.nih.gov/pubmed/37174427
http://dx.doi.org/10.3390/foods12091889
work_keys_str_mv AT xiangshuna anewduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT zhanghanyue anewduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT chaxiaoyan anewduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT linyingting anewduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT shangying anewduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT xiangshuna newduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT zhanghanyue newduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT chaxiaoyan newduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT linyingting newduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood
AT shangying newduplexrecombinasepolymeraseamplificationdrpamethodforthesimultaneousandrapiddetectionofshigellaandbacilluscereusinfood