Improved detection of Little cherry virus-2 using a hydrolysis probe to manage the Pacific Northwest little cherry disease epidemic.
Little cherry virus-2 (LChV-2) is a viral pathogen that is reaching epidemic levels in Washington state. This virus is insect-vectored and has significant impacts on sweet cherry production. To aid growers in making informed management decisions we sought to develop a diagnostic assay to better detect isolates of LChV-2 currently found in Washington, allowing for more accurate estimations of disease occurrence. This study showed that there were two distinct genotypes of LChV-2 present in Washington state. This information was used to develop an up-to-date reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) assay, which was then optimized, validated, and compared to four previously published assays using a panel of field samples. This comparison demonstrated that the newly developed assay provided greater sensitivity, accurately detecting less than 10 copies per reaction and could detect both LChV-2 genotypes. Finally, we examined the effect of potential inhibitors in various tissue types from cherry, finding that young leaf tissue affected sensitivity of detection less than root tissues.