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Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food

Salmonella can cause serious foodborne diseases. We have developed a lateral flow immunoassay combined with recombinase polymerase amplification (LFD-RPA) for detection of Salmonella in food. The conserved fragment (fimY) was selected as the target gene. Under an optimal condition (37 °C, 10 min), t...

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Autores principales: Li, Jiali, Ma, Biao, Fang, Jiehong, Zhi, Antong, Chen, Erjing, Xu, Ying, Yu, Xiaoping, Sun, Chuanxin, Zhang, Mingzhou
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022641/
https://www.ncbi.nlm.nih.gov/pubmed/31887998
http://dx.doi.org/10.3390/foods9010027
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author Li, Jiali
Ma, Biao
Fang, Jiehong
Zhi, Antong
Chen, Erjing
Xu, Ying
Yu, Xiaoping
Sun, Chuanxin
Zhang, Mingzhou
author_facet Li, Jiali
Ma, Biao
Fang, Jiehong
Zhi, Antong
Chen, Erjing
Xu, Ying
Yu, Xiaoping
Sun, Chuanxin
Zhang, Mingzhou
author_sort Li, Jiali
collection PubMed
description Salmonella can cause serious foodborne diseases. We have developed a lateral flow immunoassay combined with recombinase polymerase amplification (LFD-RPA) for detection of Salmonella in food. The conserved fragment (fimY) was selected as the target gene. Under an optimal condition (37 °C, 10 min), the sensitivity was 12 colony-forming units (CFU)/mL in a pure culture. Testing with 16 non-Salmonella strains as controls revealed that LFD-RPA was specific to the fimY gene of Salmonella. The established assay could detect Salmonella at concentrations as low as 1.29 × 10(2) CFU/mL in artificially contaminated samples. This detection was at a slightly higher level than that for a pure bacterial culture. Combined with the test strip reader, the LFD-RPA is a feasible method for quantitative detection of Salmonella based on the test line intensity, which was the ratio for the test line and control line of the reflected light. The method could be a potential point-of-care test in limited resource areas and provides a new approach and technical support for the diagnosis of food safety.
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spelling pubmed-70226412020-03-09 Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food Li, Jiali Ma, Biao Fang, Jiehong Zhi, Antong Chen, Erjing Xu, Ying Yu, Xiaoping Sun, Chuanxin Zhang, Mingzhou Foods Article Salmonella can cause serious foodborne diseases. We have developed a lateral flow immunoassay combined with recombinase polymerase amplification (LFD-RPA) for detection of Salmonella in food. The conserved fragment (fimY) was selected as the target gene. Under an optimal condition (37 °C, 10 min), the sensitivity was 12 colony-forming units (CFU)/mL in a pure culture. Testing with 16 non-Salmonella strains as controls revealed that LFD-RPA was specific to the fimY gene of Salmonella. The established assay could detect Salmonella at concentrations as low as 1.29 × 10(2) CFU/mL in artificially contaminated samples. This detection was at a slightly higher level than that for a pure bacterial culture. Combined with the test strip reader, the LFD-RPA is a feasible method for quantitative detection of Salmonella based on the test line intensity, which was the ratio for the test line and control line of the reflected light. The method could be a potential point-of-care test in limited resource areas and provides a new approach and technical support for the diagnosis of food safety. MDPI 2019-12-26 /pmc/articles/PMC7022641/ /pubmed/31887998 http://dx.doi.org/10.3390/foods9010027 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Li, Jiali
Ma, Biao
Fang, Jiehong
Zhi, Antong
Chen, Erjing
Xu, Ying
Yu, Xiaoping
Sun, Chuanxin
Zhang, Mingzhou
Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
title Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
title_full Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
title_fullStr Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
title_full_unstemmed Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
title_short Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
title_sort recombinase polymerase amplification (rpa) combined with lateral flow immunoassay for rapid detection of salmonella in food
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022641/
https://www.ncbi.nlm.nih.gov/pubmed/31887998
http://dx.doi.org/10.3390/foods9010027
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