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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...

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Autores principales: Cheng, Kewen, Pan, Daodong, Teng, Jun, Yao, Li, Ye, Yingwang, Xue, Feng, Xia, Fan, Chen, Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
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.
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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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