<abstract xmlns="http://www.w3.org/1999/xhtml">

<p>Recombinase polymerase amplification (RPA) is an isothermal <italic>in vitro</italic> 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, <italic>Meloidogyne artiellia</italic>. 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.</p>
</abstract>

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.

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

California Department of Food and Agriculture

Journal of Nematology

This work is licensed under the Creative Commons Attribution 4.0 International License.

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

Recombinase polymerase amplification (RPA) is an isothermal in vitro nucleic acid amplification technique that has been adopted for simple, robust, rapid,...

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

Article Category: Research Paper

Online veröffentlicht: 05. Juli 2024

Seitenbereich: -

Eingereicht: 12. Feb. 2024

DOI: https://doi.org/10.2478/jofnem-2024-0023

Schlüsselwörter
chickpea, diagnostics, ITS rRNA, RPA, species-specific primer

© 2024 Sergei A. Subbotin et al., published by Sciendo

This work is licensed under the Creative Commons Attribution 4.0 International License.

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

Article Category: Research Paper

Online veröffentlicht: 05. Juli 2024

Seitenbereich: -

Eingereicht: 12. Feb. 2024

DOI: https://doi.org/10.2478/jofnem-2024-0023

Schlüsselwörterchickpea, diagnostics, ITS rRNA, RPA, species-specific primer

© 2024 Sergei A. Subbotin et al., published by Sciendo

This work is licensed under the Creative Commons Attribution 4.0 International License.

Schlüsselwörter
chickpea, diagnostics, ITS rRNA, RPA, species-specific primer