One-step RT-LAMP analysis for efficient detection of Odontoglossum ringspot virus from Oncidiinae hybrids (Orchidaceae).

Odontoglossum ringspot virus (ORSV) is one of the most serious pathogens in the orchid industry. The production of virus-free orchid seedlings is the most efficient strategy to counter this virus problem. Therefore, an easy, efficient process to detect viruses is important to the orchid industry. One-step reverse transcription (RT) loop-mediated isothermal amplification (LAMP) technique was developed to detect ORSV in Oncidiinae hybrids. RT-LAMP analysis primers were designed based on the conserved regions of the RNA-dependent RNA polymerase (RdRp) gene among ORSV isolates. RT-LAMP reaction conditions were optimized to detect ORSV from Oncidiinae hybrids. This one-step RT-LAMP method proved to be easy, efficient, sensitive, and inexpensive for detecting ORSV in orchids and is the first application of a one-step RT-LAMP to detect ORSV.