Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a system

With the increase in cross-border transmission in the context of globalization, the necessity for developing rapid and accurate detection methods for plant pathogens has become critical. This study introduces a recombinase polymerase amplification (RPA) technique combined with CRISPR/Cas12a cleavage...

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Main Authors: Dorji Phurbu, Zhipeng Feng, Lei Cai, Fang Liu
Format: Article
Language:English
Published: Pensoft Publishers 2025-06-01
Series:IMA Fungus
Online Access:https://imafungus.pensoft.net/article/153604/download/pdf/
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author Dorji Phurbu
Zhipeng Feng
Lei Cai
Fang Liu
author_facet Dorji Phurbu
Zhipeng Feng
Lei Cai
Fang Liu
author_sort Dorji Phurbu
collection DOAJ
description With the increase in cross-border transmission in the context of globalization, the necessity for developing rapid and accurate detection methods for plant pathogens has become critical. This study introduces a recombinase polymerase amplification (RPA) technique combined with CRISPR/Cas12a cleavage and fluorescence-based detection systems (FRB) or paper-based lateral flow strips (PLFS) for the rapid on-site detection of invasive alien fungi, specifically Alternaria triticina and Plenodomus libanotidis, which pose significant threats to agriculture and biodiversity. The results demonstrate that either RPA-CRISPR/Cas12a-FRB or RPA-CRISPR/Cas12a-PLFS can accurately detect the target species within 30 min, with a sensitivity of up to 10 pg/μL. These portable and easy-to-use assays are suitable for rapid on-site screening of plant pathogenic fungi in plant tissues, enabling applications in disease control and port quarantine.
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institution Kabale University
issn 2210-6359
language English
publishDate 2025-06-01
publisher Pensoft Publishers
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series IMA Fungus
spelling doaj-art-a987ef3b75ce4a13a24b5000e05ad0882025-08-20T03:44:47ZengPensoft PublishersIMA Fungus2210-63592025-06-011611710.3897/imafungus.16.153604153604Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a systemDorji Phurbu0Zhipeng Feng1Lei Cai2Fang Liu3Institute of Plateau Biology of Xizang Autonomous RegionChinese Academy of SciencesUniversity of Chinese Academy of SciencesUniversity of Chinese Academy of SciencesWith the increase in cross-border transmission in the context of globalization, the necessity for developing rapid and accurate detection methods for plant pathogens has become critical. This study introduces a recombinase polymerase amplification (RPA) technique combined with CRISPR/Cas12a cleavage and fluorescence-based detection systems (FRB) or paper-based lateral flow strips (PLFS) for the rapid on-site detection of invasive alien fungi, specifically Alternaria triticina and Plenodomus libanotidis, which pose significant threats to agriculture and biodiversity. The results demonstrate that either RPA-CRISPR/Cas12a-FRB or RPA-CRISPR/Cas12a-PLFS can accurately detect the target species within 30 min, with a sensitivity of up to 10 pg/μL. These portable and easy-to-use assays are suitable for rapid on-site screening of plant pathogenic fungi in plant tissues, enabling applications in disease control and port quarantine.https://imafungus.pensoft.net/article/153604/download/pdf/
spellingShingle Dorji Phurbu
Zhipeng Feng
Lei Cai
Fang Liu
Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a system
IMA Fungus
title Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a system
title_full Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a system
title_fullStr Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a system
title_full_unstemmed Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a system
title_short Rapid and sensitive diagnosis of plant quarantine fungi Alternaria triticina and Plenodomus libanotidis based on the RPA-CRISPR/Cas12a system
title_sort rapid and sensitive diagnosis of plant quarantine fungi alternaria triticina and plenodomus libanotidis based on the rpa crispr cas12a system
url https://imafungus.pensoft.net/article/153604/download/pdf/
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