Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid and Sensitive Detection of Barley Yellow Dwarf Virus in Oat

Rapid and Visual Detection of Barley Yellow Dwarf Virus by Reverse Transcription Recombinase Polymerase Amplification with Lateral Flow Strips

“…RT-PCR products (approximately 595 bp) for BYDV detection were visualized on 1.5% agarose gels. An actin gene from Rhopalosiphum padi was used as an internal control ( Kim et al, 2020 ). RT-RPA-LFS, reverse transcription recombinase polymerase amplification combined with a lateral flow strips; BYDV, barley yellow dwarf virus; RT-PCR, reverse transcription polymerase chain reaction.…”

“…In general, RPA products can be analyzed using agarose gel electrophoresis, real-time systems with probe-based fluorescence, or lateral flow strips (LFS). Recently, an RPA assay was developed to detect BYDV in oat using an agarose gel electrophoresis system ( Kim et al, 2020 ). RPA system combined with LFS (termed RPA-LFS) requires a set of primers, a 5′-biotin-labeled primer and its complementary sequence and a probe that is typically labeled with a FAM (carboxyfluorescein) antigen at the 5′ terminus, a tetradrofuran (THF) spacer at the center, and a C3 spacer at the 3′ terminus.…”

“…In addition, the RT-RPA-LFS assay was validated with BYDV-infected field samples and aphids. In a previous study, a conventional RT-RPA assay was developed for the detection of BYDV in oat plants ( Kim et al, 2020 ); however, this assay requires more time for BYDV detection because it was developed for agarose gel-based electrophoresis detection systems. Moreover, RT-LAMP assay, an isothermal nucleic acid amplification method, was developed for the detection of BYDV in oat plants ( Zhao et al, 2010 ); however, it requires six primers, causing difficulty regarding primer design from the BYDV genome and is performed at 65 °C for 80 min.…”

“…In general, RPA products can be analyzed using agarose gel electrophoresis, real-time systems with probebased fluorescence, or lateral flow strips (LFS). Recently, an RPA assay was developed to detect BYDV in oat using an agarose gel electrophoresis system (Kim et al, 2020).…”

Characterization and Establishment of a Recombinase Polymerase Amplification Assay for Rapid Detection of Tomato Spotted Wilt Virus in Pepper

“…In recent years, RPA has been developed rapidly in the detection of plant viruses [21]. Since the emergence of application of RPA to detect little cherry virus 2 (LChV2) [22], more and more plant viruses have been detected by RPA, especially in 2020, such as chilli veinal mottle virus (ChiVMV), milk vetch dwarf virus (MDV), piper yellow mottle virus (PYMoV), cardamom vein clearing virus (CdVCV), barley yellow dwarf virus (BYDV), cucurbit yellow stunting disorder virus (CYSDV) [23][24][25][26][27][28]. In this study, pepper leaves with suspected symptoms of TSWV were collected from Tieling city of Liaoning Province in 2020.…”