Development of reverse transcription recombinase polymerase amplification assay for rapid diagnostics of Peanut mottle virus.

Peanut mottle virus (PeMoV) is a single-stranded RNA virus transmitted through seeds and aphids that affects peanut crops worldwide. Currently, Enzyme Linked Immune-Sorbent assays and Reverse-Transcription Polymerase Chain Reaction techniques are widely employed to detect PeMoV in infected plants. ELISA is labor-intensive and time-consuming, as it involves the preparation of buffers and the production of polyclonal antibodies. Even though RT-PCR bypasses the need for buffer preparation and antibody production, it demands trained professional's manpower, requires expensive equipment like thermal cyclers, and involves complex procedures such as RNA isolation and cDNA conversion. To avoid these constraints, there is a need for a fast, reliable, efficient, and economical method for detecting PeMoV to ensure the production of healthy seeds. This study optimized the Reverse Transcriptase Recombinase Polymerase Amplification (RT-RPA) method by eliminating the steps of RNA extraction, cDNA conversion, and the use of a thermal cycler. The optimized RT-RPA assay successfully detected PeMoV at concentrations as low as 10 Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01545-3.