Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissues
Abstract Meloidogyne enterolobii, a guava root-knot nematode, is a highly virulent pest in tropical and subtropical regions causing galls or knots in roots of diverse plant species posing a serious threat to agriculture. Managing this nematode is challenging due to limitations in conventional identi...
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Nature Portfolio
2025-01-01
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| Online Access: | https://doi.org/10.1038/s41598-024-83214-9 |
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| author | Arunee Suwanngam Philipp H. Schiffer Anongnuch Sasnarukkit Suradej Siripattanapipong Kansiree Jindapunnapat Buncha Chinnasri Toon Ruang-areerate |
| author_facet | Arunee Suwanngam Philipp H. Schiffer Anongnuch Sasnarukkit Suradej Siripattanapipong Kansiree Jindapunnapat Buncha Chinnasri Toon Ruang-areerate |
| author_sort | Arunee Suwanngam |
| collection | DOAJ |
| description | Abstract Meloidogyne enterolobii, a guava root-knot nematode, is a highly virulent pest in tropical and subtropical regions causing galls or knots in roots of diverse plant species posing a serious threat to agriculture. Managing this nematode is challenging due to limitations in conventional identification based on isolation and microscopic classification requiring expertise and time. A colorimetric and fluorescent LAMP assay using simplified extraction method targeting rDNA-ITS region was developed to detect M. enterolobii DNA. The Men-LAMP assay exhibits simple procedure and achievable outcomes directly from root gall samples within 75 to 80 min, using a simplified Worm Lysis Buffer Plus (WLB +) extraction and the LAMP assay. The results could be interpreted using color and fluorescence without requiring post-amplification to minimize any possibility of contamination. The specificity showed no cross amplification with other plant-parasitic nematodes, a sensitivity was limited to 2.89 ng/μL. Our study proposes a sensitive, specific and time-efficient diagnostic tool for M. enterolobii infection as an alternative promising method for rapid and effective diagnosis at point-of-service to manage and control of M. enterolobii in export plants that can contribute to the degradation of trade restrictions and streamline of the international quarantine inspection process. |
| format | Article |
| id | doaj-art-2d89be4989724cb58e68c6c1d0ce7ca4 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-2d89be4989724cb58e68c6c1d0ce7ca42025-01-05T12:23:15ZengNature PortfolioScientific Reports2045-23222025-01-0115111210.1038/s41598-024-83214-9Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissuesArunee Suwanngam0Philipp H. Schiffer1Anongnuch Sasnarukkit2Suradej Siripattanapipong3Kansiree Jindapunnapat4Buncha Chinnasri5Toon Ruang-areerate6Tropical Agriculture International Program, Faculty of Agriculture, Kasetsart UniversityInstitute for Zoology, University of CologneDepartment of Plant Pathology, Faculty of Agriculture, Kasetsart UniversityDepartment of Microbiology, Faculty of Science, Mahidol UniversityDepartment of Entomology and Plant Pathology, Faculty of Agriculture, Khon Kaen UniversityDepartment of Plant Pathology, Faculty of Agriculture, Kasetsart UniversityDepartment of Parasitology, Phramongkutklao College of MedicineAbstract Meloidogyne enterolobii, a guava root-knot nematode, is a highly virulent pest in tropical and subtropical regions causing galls or knots in roots of diverse plant species posing a serious threat to agriculture. Managing this nematode is challenging due to limitations in conventional identification based on isolation and microscopic classification requiring expertise and time. A colorimetric and fluorescent LAMP assay using simplified extraction method targeting rDNA-ITS region was developed to detect M. enterolobii DNA. The Men-LAMP assay exhibits simple procedure and achievable outcomes directly from root gall samples within 75 to 80 min, using a simplified Worm Lysis Buffer Plus (WLB +) extraction and the LAMP assay. The results could be interpreted using color and fluorescence without requiring post-amplification to minimize any possibility of contamination. The specificity showed no cross amplification with other plant-parasitic nematodes, a sensitivity was limited to 2.89 ng/μL. Our study proposes a sensitive, specific and time-efficient diagnostic tool for M. enterolobii infection as an alternative promising method for rapid and effective diagnosis at point-of-service to manage and control of M. enterolobii in export plants that can contribute to the degradation of trade restrictions and streamline of the international quarantine inspection process.https://doi.org/10.1038/s41598-024-83214-9Meloidogyne enterolobiiLAMPExtractionSYBRRootParasite |
| spellingShingle | Arunee Suwanngam Philipp H. Schiffer Anongnuch Sasnarukkit Suradej Siripattanapipong Kansiree Jindapunnapat Buncha Chinnasri Toon Ruang-areerate Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissues Scientific Reports Meloidogyne enterolobii LAMP Extraction SYBR Root Parasite |
| title | Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissues |
| title_full | Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissues |
| title_fullStr | Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissues |
| title_full_unstemmed | Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissues |
| title_short | Development of colorimetric and fluorescent closed tube LAMP assay using simplified extraction for diagnosis of Meloidogyne enterolobii in root tissues |
| title_sort | development of colorimetric and fluorescent closed tube lamp assay using simplified extraction for diagnosis of meloidogyne enterolobii in root tissues |
| topic | Meloidogyne enterolobii LAMP Extraction SYBR Root Parasite |
| url | https://doi.org/10.1038/s41598-024-83214-9 |
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