Development of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood disease

Efficient and accurate detection of Ralstonia syzygii subsp. celebesensis (Rsc), the pathogen responsible for banana blood disease, is vital for controlling its detrimental effects on banana cultivation. This study developed and assessed a loopmediated isothermal amplification (LAMP), which detected...

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Main Authors: Kaennuea Thipsukon, Thanyasiriwat Thanwanit, Rattanapolsan Ladawan, Ponpang-Nga Pancheewan, Sangdee Aphidech, Kawicha Praphat
Format: Article
Language:English
Published: University of Belgrade, University of Novi Sad 2025-01-01
Series:Archives of Biological Sciences
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Online Access:https://doiserbia.nb.rs/img/doi/0354-4664/2025/0354-46642500013K.pdf
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author Kaennuea Thipsukon
Thanyasiriwat Thanwanit
Rattanapolsan Ladawan
Ponpang-Nga Pancheewan
Sangdee Aphidech
Kawicha Praphat
author_facet Kaennuea Thipsukon
Thanyasiriwat Thanwanit
Rattanapolsan Ladawan
Ponpang-Nga Pancheewan
Sangdee Aphidech
Kawicha Praphat
author_sort Kaennuea Thipsukon
collection DOAJ
description Efficient and accurate detection of Ralstonia syzygii subsp. celebesensis (Rsc), the pathogen responsible for banana blood disease, is vital for controlling its detrimental effects on banana cultivation. This study developed and assessed a loopmediated isothermal amplification (LAMP), which detected as few as 10 cells per reaction, significantly surpassing conventional PCR, which required a minimum of 10³ cells per reaction. The LAMP assay utilizes real-time fluorescence-based Ct values and Tm analysis, ensuring reliable detection by confirming target specificity. The assay effectively identified Rsc in Rsc-spiked banana sucker samples and naturally infected field specimens using a simple, purification-free boiling DNA extraction method, making it highly adaptable for field diagnostics. By integrating real-time fluorescence detection with Tm validation, this study presents an advanced, field-deployable molecular diagnostic tool with superior sensitivity and specificity, addressing critical limitations of existing Rsc detection methods and enhancing early disease surveillance in banana cultivation.
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institution Kabale University
issn 0354-4664
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language English
publishDate 2025-01-01
publisher University of Belgrade, University of Novi Sad
record_format Article
series Archives of Biological Sciences
spelling doaj-art-e4dacc9f06e34f5590ce40e467beef802025-08-20T03:58:44ZengUniversity of Belgrade, University of Novi SadArchives of Biological Sciences0354-46641821-43392025-01-0177215917110.2298/ABS250310013K0354-46642500013KDevelopment of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood diseaseKaennuea Thipsukon0https://orcid.org/0009-0005-5144-5258Thanyasiriwat Thanwanit1https://orcid.org/0000-0002-0234-120XRattanapolsan Ladawan2https://orcid.org/0009-0009-0422-1189Ponpang-Nga Pancheewan3https://orcid.org/0000-0002-9465-0316Sangdee Aphidech4https://orcid.org/0000-0003-4401-8985Kawicha Praphat5https://orcid.org/0000-0003-4230-2720Plant Genome and Disease Research Unit, Department of Agriculture and Resources, Faculty of Natural Resources and Agro-Industry, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, ThailandPlant Genome and Disease Research Unit, Department of Agriculture and Resources, Faculty of Natural Resources and Agro-Industry, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, ThailandPlant Genome and Disease Research Unit, Department of Agriculture and Resources, Faculty of Natural Resources and Agro-Industry, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, ThailandPlant Genome and Disease Research Unit, Department of Agriculture and Resources, Faculty of Natural Resources and Agro-Industry, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, ThailandDepartment of Biology, Faculty of Science, Mahasarakham University, Kantarawichai District, Maha Sarakham, Thailand + Microbiology and Applied Microbiology Research Unit, Faculty of Science, Mahasarakham University, Kantarawichai District, Maha Sarakham, ThailandPlant Genome and Disease Research Unit, Department of Agriculture and Resources, Faculty of Natural Resources and Agro-Industry, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, ThailandEfficient and accurate detection of Ralstonia syzygii subsp. celebesensis (Rsc), the pathogen responsible for banana blood disease, is vital for controlling its detrimental effects on banana cultivation. This study developed and assessed a loopmediated isothermal amplification (LAMP), which detected as few as 10 cells per reaction, significantly surpassing conventional PCR, which required a minimum of 10³ cells per reaction. The LAMP assay utilizes real-time fluorescence-based Ct values and Tm analysis, ensuring reliable detection by confirming target specificity. The assay effectively identified Rsc in Rsc-spiked banana sucker samples and naturally infected field specimens using a simple, purification-free boiling DNA extraction method, making it highly adaptable for field diagnostics. By integrating real-time fluorescence detection with Tm validation, this study presents an advanced, field-deployable molecular diagnostic tool with superior sensitivity and specificity, addressing critical limitations of existing Rsc detection methods and enhancing early disease surveillance in banana cultivation.https://doiserbia.nb.rs/img/doi/0354-4664/2025/0354-46642500013K.pdfbanana blood diseasedna fingerprintinglamp assayralstonia syzygii subsp. celebesensisrepetitive-pcr
spellingShingle Kaennuea Thipsukon
Thanyasiriwat Thanwanit
Rattanapolsan Ladawan
Ponpang-Nga Pancheewan
Sangdee Aphidech
Kawicha Praphat
Development of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood disease
Archives of Biological Sciences
banana blood disease
dna fingerprinting
lamp assay
ralstonia syzygii subsp. celebesensis
repetitive-pcr
title Development of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood disease
title_full Development of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood disease
title_fullStr Development of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood disease
title_full_unstemmed Development of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood disease
title_short Development of an updated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Ralstonia syzygii subsp. celebesensis (Rsc), the causative agent of banana blood disease
title_sort development of an updated loop mediated isothermal amplification lamp assay for the rapid detection of ralstonia syzygii subsp celebesensis rsc the causative agent of banana blood disease
topic banana blood disease
dna fingerprinting
lamp assay
ralstonia syzygii subsp. celebesensis
repetitive-pcr
url https://doiserbia.nb.rs/img/doi/0354-4664/2025/0354-46642500013K.pdf
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