Immunocapture-Multiplex RT-PCR for the Simultaneous Detection and Identification of Plant Viruses and Their Strains: Study Case， Potato Virus Y (PVY).

Immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) is a sensitive， reproducible， and robust method for the detection and identification of RNA viruses. The IC step provides a simple method to isolate virus particles from plant tissue， particularly when inhibitory substances are present， and thus enables subsequent use of RT-PCR amplification for large-scale virus testing and typing. The multiplex format of the PCR is often used for the detection and identification of multiple virus/strain simultaneously to save time， labor， and cost. Potato virus Y (PVY) is one of the most economically important viruses infecting potato worldwide. PVY exists as a complex of at least nine strains and many more unclassified recombinants that vary in their genome structures， phenotypes， and their economic importance. In the current chapter， a detailed protocol of an IC-based， multiplex RT-PCR assay for the detection and identification of various PVY strains is described.