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Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay

Phytophthora cactorum is a devastating pathogen that infects a wide range of plants and causes Phytophthora rot disease, which has resulted in great economic losses in crop production. Therefore, the rapid and practicable detection of P. cactorum is important for disease monitoring and forecasting....

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Autores principales: Lu, Xinyu, Xu, Heng, Song, Wen, Yang, Zitong, Yu, Jia, Tian, Yuee, Jiang, Min, Shen, Danyu, Dou, Daolong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8189726/
https://www.ncbi.nlm.nih.gov/pubmed/34127941
http://dx.doi.org/10.1186/s42483-021-00089-8
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author Lu, Xinyu
Xu, Heng
Song, Wen
Yang, Zitong
Yu, Jia
Tian, Yuee
Jiang, Min
Shen, Danyu
Dou, Daolong
author_facet Lu, Xinyu
Xu, Heng
Song, Wen
Yang, Zitong
Yu, Jia
Tian, Yuee
Jiang, Min
Shen, Danyu
Dou, Daolong
author_sort Lu, Xinyu
collection PubMed
description Phytophthora cactorum is a devastating pathogen that infects a wide range of plants and causes Phytophthora rot disease, which has resulted in great economic losses in crop production. Therefore, the rapid and practicable detection of P. cactorum is important for disease monitoring and forecasting. In this study, we developed a lateral flow recombinase polymerase amplification (LF-RPA) assay for the sensitive visual detection of P. cactorum. Specific primers for P. cactorum were designed based on the ras-related protein gene Ypt1; all 10 P. cactorum isolates yielded positive detection results, whereas no cross-reaction occurred in related oomycete or fungal species. The detection limit for the LF-RPA assay was 100 fg of genomic DNA under optimized conditions. Combined with a simplified alkaline lysis method for plant DNA extraction, the LF-RPA assay successfully detected P. cactorum in naturally diseased strawberry samples without specialized equipment within 40 min. Thus, the LF-RPA assay developed in this study is a rapid, simple, and accurate method for the detection of P. cactorum, with the potential for further application in resource-limited laboratories. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s42483-021-00089-8.
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spelling pubmed-81897262021-06-10 Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay Lu, Xinyu Xu, Heng Song, Wen Yang, Zitong Yu, Jia Tian, Yuee Jiang, Min Shen, Danyu Dou, Daolong Phytopathol Res Research Phytophthora cactorum is a devastating pathogen that infects a wide range of plants and causes Phytophthora rot disease, which has resulted in great economic losses in crop production. Therefore, the rapid and practicable detection of P. cactorum is important for disease monitoring and forecasting. In this study, we developed a lateral flow recombinase polymerase amplification (LF-RPA) assay for the sensitive visual detection of P. cactorum. Specific primers for P. cactorum were designed based on the ras-related protein gene Ypt1; all 10 P. cactorum isolates yielded positive detection results, whereas no cross-reaction occurred in related oomycete or fungal species. The detection limit for the LF-RPA assay was 100 fg of genomic DNA under optimized conditions. Combined with a simplified alkaline lysis method for plant DNA extraction, the LF-RPA assay successfully detected P. cactorum in naturally diseased strawberry samples without specialized equipment within 40 min. Thus, the LF-RPA assay developed in this study is a rapid, simple, and accurate method for the detection of P. cactorum, with the potential for further application in resource-limited laboratories. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s42483-021-00089-8. BioMed Central 2021-06-10 2021 /pmc/articles/PMC8189726/ /pubmed/34127941 http://dx.doi.org/10.1186/s42483-021-00089-8 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research
Lu, Xinyu
Xu, Heng
Song, Wen
Yang, Zitong
Yu, Jia
Tian, Yuee
Jiang, Min
Shen, Danyu
Dou, Daolong
Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay
title Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay
title_full Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay
title_fullStr Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay
title_full_unstemmed Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay
title_short Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay
title_sort rapid and simple detection of phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8189726/
https://www.ncbi.nlm.nih.gov/pubmed/34127941
http://dx.doi.org/10.1186/s42483-021-00089-8
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