One-step multiplex DiRT-PCR for PLRV, PVY, PVM, PVS, PVA and PVX ready for routine testing directly on tuber sap

Abstract Potato viruses PLRV, PVY, PVM, PVA, PVX and PVS can cause up to 90% loss of potato harvest. Therefore, they are monitored by law in many countries using DAS-ELISA or conventional real-time RT-qPCR. Previously, we developed a multiplex DiRT-PCR (Direct reverse transcript – polymerase chain reaction), which works directly on diluted tuber sap thus saves time chemical processing for RNA extraction space the glasshouse. So far, this method only ran single tubers is not practical routine testing. We now able sensitively test presence six two reactions pooled samples ten tubers. Here show that there an “almost perfect” agreement (Gwet’s AC1 index) comparing with commercial RT-qPCR kit rapid method. The ready be used