Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia

Recombinase polymerase amplification (RPA) is an isothermal 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...

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Bibliographic Details
Published in:Journal of nematology 2024-03, Vol.56 (1), p.20240023-1523
Main Authors: Subbotin, Sergei A., Palomares-Rius, Juan E., Castillo, Pablo
Format: Article
Language:English
Subjects:
chickpea
diagnostics
ITS rRNA
Research Paper
RPA
species-specific primer
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Summary:Recombinase polymerase amplification (RPA) is an isothermal 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, . 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.
ISSN:0022-300X
2640-396X
2640-396X
DOI:10.2478/jofnem-2024-0023