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A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification

Foodborne pathogenic bacteria have become a worldwide threat to human health, and rapid and sensitive bacterial detection methods are urgently needed. In this study, a facile microfluidic chip was developed and combined with recombinase-aided amplification (RAA) for rapid and sensitive detection of...

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Autores principales: Wu, Shangyi, Duan, Hong, Zhang, Yingchao, Wang, Siyuan, Zheng, Lingyan, Cai, Gaozhe, Lin, Jianhan, Yue, Xiqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9496460/
https://www.ncbi.nlm.nih.gov/pubmed/36140111
http://dx.doi.org/10.3390/bios12090726
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author Wu, Shangyi
Duan, Hong
Zhang, Yingchao
Wang, Siyuan
Zheng, Lingyan
Cai, Gaozhe
Lin, Jianhan
Yue, Xiqing
author_facet Wu, Shangyi
Duan, Hong
Zhang, Yingchao
Wang, Siyuan
Zheng, Lingyan
Cai, Gaozhe
Lin, Jianhan
Yue, Xiqing
author_sort Wu, Shangyi
collection PubMed
description Foodborne pathogenic bacteria have become a worldwide threat to human health, and rapid and sensitive bacterial detection methods are urgently needed. In this study, a facile microfluidic chip was developed and combined with recombinase-aided amplification (RAA) for rapid and sensitive detection of Salmonella typhimurium using a non-contact eddy heater for dynamic lysis of bacterial cells and a 3D-printed fan-shaped active mixer for continuous-flow mixing. First, the bacterial sample was injected into the chip to flow through the spiral channel coiling around an iron rod under an alternating electromagnetic field, resulting in the dynamic lysis of bacterial cells by this non-contact eddy heater to release their nucleic acids. After cooling to ~75 °C, these nucleic acids were continuous-flow mixed with magnetic silica beads using the fan-shaped mixer and captured in the separation chamber using a magnet. Finally, the captured nucleic acids were eluted by the eluent from the beads to flow into the detection chamber, followed by RAA detection of nucleic acids to determine the bacterial amount. Under the optimal conditions, this microfluidic chip was able to quantitatively detect Salmonella typhimurium from 1.1 × 10(2) to 1.1 × 10(5) CFU/mL in 40 min with a detection limit of 89 CFU/mL and might be prospective to offer a simple, low-cost, fast and specific bacterial detection technique for ensuring food safety.
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spelling pubmed-94964602022-09-23 A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification Wu, Shangyi Duan, Hong Zhang, Yingchao Wang, Siyuan Zheng, Lingyan Cai, Gaozhe Lin, Jianhan Yue, Xiqing Biosensors (Basel) Article Foodborne pathogenic bacteria have become a worldwide threat to human health, and rapid and sensitive bacterial detection methods are urgently needed. In this study, a facile microfluidic chip was developed and combined with recombinase-aided amplification (RAA) for rapid and sensitive detection of Salmonella typhimurium using a non-contact eddy heater for dynamic lysis of bacterial cells and a 3D-printed fan-shaped active mixer for continuous-flow mixing. First, the bacterial sample was injected into the chip to flow through the spiral channel coiling around an iron rod under an alternating electromagnetic field, resulting in the dynamic lysis of bacterial cells by this non-contact eddy heater to release their nucleic acids. After cooling to ~75 °C, these nucleic acids were continuous-flow mixed with magnetic silica beads using the fan-shaped mixer and captured in the separation chamber using a magnet. Finally, the captured nucleic acids were eluted by the eluent from the beads to flow into the detection chamber, followed by RAA detection of nucleic acids to determine the bacterial amount. Under the optimal conditions, this microfluidic chip was able to quantitatively detect Salmonella typhimurium from 1.1 × 10(2) to 1.1 × 10(5) CFU/mL in 40 min with a detection limit of 89 CFU/mL and might be prospective to offer a simple, low-cost, fast and specific bacterial detection technique for ensuring food safety. MDPI 2022-09-05 /pmc/articles/PMC9496460/ /pubmed/36140111 http://dx.doi.org/10.3390/bios12090726 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
Wu, Shangyi
Duan, Hong
Zhang, Yingchao
Wang, Siyuan
Zheng, Lingyan
Cai, Gaozhe
Lin, Jianhan
Yue, Xiqing
A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification
title A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification
title_full A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification
title_fullStr A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification
title_full_unstemmed A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification
title_short A Salmonella Microfluidic Chip Combining Non-Contact Eddy Heater and 3D Fan-Shaped Mixer with Recombinase Aided Amplification
title_sort salmonella microfluidic chip combining non-contact eddy heater and 3d fan-shaped mixer with recombinase aided amplification
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9496460/
https://www.ncbi.nlm.nih.gov/pubmed/36140111
http://dx.doi.org/10.3390/bios12090726
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