Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia
Recombinase polymerase amplification (RPA) is an isothermal in vitro nucleic acid amplification technique that has been adopted for simple, robust, rapid, reliable diagnostics of nematodes. In this study, the real-time RPA assay and RPA assay combined with lateral flow dipsticks (LF-RPA) have been d...
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| Format: | Article |
| Language: | English |
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Sciendo
2024-07-01
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| Series: | Journal of Nematology |
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| Online Access: | https://doi.org/10.2478/jofnem-2024-0023 |
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| author | Subbotin Sergei A. Palomares-Rius Juan E. Castillo Pablo |
| author_facet | Subbotin Sergei A. Palomares-Rius Juan E. Castillo Pablo |
| author_sort | Subbotin Sergei A. |
| collection | DOAJ |
| description | Recombinase polymerase amplification (RPA) is an isothermal in vitro nucleic acid amplification technique that has been adopted for simple, robust, rapid, reliable diagnostics of nematodes. In this study, the real-time RPA assay and RPA assay combined with lateral flow dipsticks (LF-RPA) have been developed targeting the ITS rRNA gene of the British root-knot nematode, Meloidogyne artiellia. The assay provided specific and rapid detection of this root-knot nematode species from crude nematode extracts without a DNA extraction step with a sensitivity of 0.125 second-stage juvenile (J2) specimen per a reaction tube for real-time RPA during 11 min and a sensitivity of 0.5 J2 specimens per a reaction tube for LF-RPA during 25 min. The RPA assays were validated with a wide range of non-target root-knot nematodes. The LF-RPA assay has great potential for nematode diagnostics in the laboratory having minimal available equipment. |
| format | Article |
| id | doaj-art-db93a5e4c2df4ccaa721ec7fc8543413 |
| institution | OA Journals |
| issn | 2640-396X |
| language | English |
| publishDate | 2024-07-01 |
| publisher | Sciendo |
| record_format | Article |
| series | Journal of Nematology |
| spelling | doaj-art-db93a5e4c2df4ccaa721ec7fc85434132025-08-20T02:23:39ZengSciendoJournal of Nematology2640-396X2024-07-0156111788152310.2478/jofnem-2024-0023Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artielliaSubbotin Sergei A.0Palomares-Rius Juan E.1Castillo Pablo2Plant Pest Diagnostic Center, California Department of Food and Agriculture, 3294 Meadowview Road, 95832, Sacramento, CA, USAInstituto de Agricultura Sostenible (IAS), Consejo Superior de Investigaciones Científicas (CSIC), Campus de Excelencia Internacional Agroalimentario, ceiA3, Avda. Menendez Pidal s/n, 14004-Córdoba, SpainInstituto de Agricultura Sostenible (IAS), Consejo Superior de Investigaciones Científicas (CSIC), Campus de Excelencia Internacional Agroalimentario, ceiA3, Avda. Menendez Pidal s/n, 14004-Córdoba, SpainRecombinase polymerase amplification (RPA) is an isothermal in vitro nucleic acid amplification technique that has been adopted for simple, robust, rapid, reliable diagnostics of nematodes. In this study, the real-time RPA assay and RPA assay combined with lateral flow dipsticks (LF-RPA) have been developed targeting the ITS rRNA gene of the British root-knot nematode, Meloidogyne artiellia. The assay provided specific and rapid detection of this root-knot nematode species from crude nematode extracts without a DNA extraction step with a sensitivity of 0.125 second-stage juvenile (J2) specimen per a reaction tube for real-time RPA during 11 min and a sensitivity of 0.5 J2 specimens per a reaction tube for LF-RPA during 25 min. The RPA assays were validated with a wide range of non-target root-knot nematodes. The LF-RPA assay has great potential for nematode diagnostics in the laboratory having minimal available equipment.https://doi.org/10.2478/jofnem-2024-0023chickpeadiagnosticsits rrnarpaspecies-specific primer |
| spellingShingle | Subbotin Sergei A. Palomares-Rius Juan E. Castillo Pablo Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia Journal of Nematology chickpea diagnostics its rrna rpa species-specific primer |
| title | Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia |
| title_full | Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia |
| title_fullStr | Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia |
| title_full_unstemmed | Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia |
| title_short | Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia |
| title_sort | recombinase polymerase amplification assay for detection of the british root knot nematode meloidogyne artiellia |
| topic | chickpea diagnostics its rrna rpa species-specific primer |
| url | https://doi.org/10.2478/jofnem-2024-0023 |
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