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Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus
Rapid detection of pathogens is of great significance for food safety and disease diagnosis. A new colorimetric method for rapid and easy detection of Vibrio parahaemolyticus (V. parahaemolyticus or Vp) has been developed in this research. A specific sequence was designed and integrated with the for...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5087389/ https://www.ncbi.nlm.nih.gov/pubmed/27690041 http://dx.doi.org/10.3390/s16101600 |
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author | Cheng, Kewen Pan, Daodong Teng, Jun Yao, Li Ye, Yingwang Xue, Feng Xia, Fan Chen, Wei |
author_facet | Cheng, Kewen Pan, Daodong Teng, Jun Yao, Li Ye, Yingwang Xue, Feng Xia, Fan Chen, Wei |
author_sort | Cheng, Kewen |
collection | PubMed |
description | Rapid detection of pathogens is of great significance for food safety and disease diagnosis. A new colorimetric method for rapid and easy detection of Vibrio parahaemolyticus (V. parahaemolyticus or Vp) has been developed in this research. A specific sequence was designed and integrated with the forward primer for molecular detection of Vp. This specific sequence was tested and treated as the horseradish peroxidase (HRP)-mimicking DNAzyme and could be amplified during the polymerase chain reaction (PCR) process. The products of PCR including the sequence of HRP-mimicking DNAzyme could produce the distinguished color in the presence of catalysis substrates. The optical signal of the catalysis reaction, which is in a linear relationship with the initial template of Vp, could be determined with the naked eye or measured with Ultraviolet-visible (UV-vis) for qualitative and quantitative detections, respectively. Based on the optical signal intensity, rapid and easy detection of Vp was successfully achieved with satisfied sensitivity and specificity. Furthermore, the detection of tdh, trh, tlh and toxR virulence genes of two Vp species (Vp 33847 and Vp 17802) were all performed successfully with this developed colorimetric integrated PCR protocol, which demonstrated potential applicability for the rapid detection of other bacteria. |
format | Online Article Text |
id | pubmed-5087389 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-50873892016-11-07 Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus Cheng, Kewen Pan, Daodong Teng, Jun Yao, Li Ye, Yingwang Xue, Feng Xia, Fan Chen, Wei Sensors (Basel) Article Rapid detection of pathogens is of great significance for food safety and disease diagnosis. A new colorimetric method for rapid and easy detection of Vibrio parahaemolyticus (V. parahaemolyticus or Vp) has been developed in this research. A specific sequence was designed and integrated with the forward primer for molecular detection of Vp. This specific sequence was tested and treated as the horseradish peroxidase (HRP)-mimicking DNAzyme and could be amplified during the polymerase chain reaction (PCR) process. The products of PCR including the sequence of HRP-mimicking DNAzyme could produce the distinguished color in the presence of catalysis substrates. The optical signal of the catalysis reaction, which is in a linear relationship with the initial template of Vp, could be determined with the naked eye or measured with Ultraviolet-visible (UV-vis) for qualitative and quantitative detections, respectively. Based on the optical signal intensity, rapid and easy detection of Vp was successfully achieved with satisfied sensitivity and specificity. Furthermore, the detection of tdh, trh, tlh and toxR virulence genes of two Vp species (Vp 33847 and Vp 17802) were all performed successfully with this developed colorimetric integrated PCR protocol, which demonstrated potential applicability for the rapid detection of other bacteria. MDPI 2016-09-28 /pmc/articles/PMC5087389/ /pubmed/27690041 http://dx.doi.org/10.3390/s16101600 Text en © 2016 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 Cheng, Kewen Pan, Daodong Teng, Jun Yao, Li Ye, Yingwang Xue, Feng Xia, Fan Chen, Wei Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus |
title | Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus |
title_full | Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus |
title_fullStr | Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus |
title_full_unstemmed | Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus |
title_short | Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus |
title_sort | colorimetric integrated pcr protocol for rapid detection of vibrio parahaemolyticus |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5087389/ https://www.ncbi.nlm.nih.gov/pubmed/27690041 http://dx.doi.org/10.3390/s16101600 |
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