title: Immunocapture-Multiplex RT-PCR for the Simultaneous Detection and Identification of Plant Viruses and Their Strains: Study Case, Potato Virus Y (PVY).
abstract: 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.
authors.affiliation: Department of Plant Soil and Entomological Sciences, University of Idaho, 875 Perimeter Drive MS 2339, Moscow, ID, 83844-2339, USA, mchikhali@uidaho.edu.
authors.forename: Mohamad
authors.initials: M
authors.lastname: Chikh-Ali
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Karasev
mesh_terms: D000914:Antibodies, Viral / Q000276:immunology*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000821:virology; D035281:Plant Tubers / Q000821:virology; D017800:Potyvirus / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D011198:Solanum tuberosum / Q000821:virology*
chemical_list: D000914:Antibodies, Viral; D012367:RNA, Viral