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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| Format: | Article |
| Language: | English |
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University of Belgrade, University of Novi Sad
2025-01-01
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| 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. |
| format | Article |
| id | doaj-art-e4dacc9f06e34f5590ce40e467beef80 |
| institution | Kabale University |
| issn | 0354-4664 1821-4339 |
| 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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