An ultra-sensitive and highly specific detection method for a newly discovered Oncotshavirus using one-step reverse transcription droplet-based digital PCR

Digital PCR is an improved detection method compared with conventional PCR. An ultra-sensitive and specific detection method for a newly discovered Oncotshavirus was established by using droplet-based digital PCR technology in this study. The detection system was highly specific and could detect the target virus in the presence of other aquatic viruses. Moreover, the limit of quantitation (LOQ) and the limit of detection (LOD) of the method were comprehensively evaluated based on the serial dilution of original virus samples and were four and one particle per reaction volume respectively. This was a substantial improvement over conventional real-time PCR. Our detection system achieved accurate and stable detection results in a variety of food matrices. Detection of the newly discovered Oncotshavirus based on droplet-based digital PCR was adaptable and applicable for routine testing.