title: Improvement of Potato virus Y (PVY) detection and quantitation using PVY(N)- and PVY(O)-specific real-time RT-PCR assays.
abstract: A Potato virus Y (PVY) single nucleotide polymorphism (A/G(2213)), recently identified as a molecular determinant of the tobacco leaf necrosis symptom induced by PVY(N) isolates, has been used as a target to develop two PVY group-specific (PVY(N) and PVY(O)) fluorescent (TaqMan-based) real-time RT-PCR assays. These procedures allow detection, characterisation, and quantitation of a wide range of PVY isolates in samples containing 10(3)-10(8) viral transcripts. Moreover, the high specificity of these two new assays make the simultaneous detection and the reliable quantitation of PVY(N) and PVY(O) isolates in mixed solutions, regardless of the Y(N)/Y(O) ratio, feasible. The high sensitivity (threshold of 10(3) copies per reaction) and the PVY group specificity of these two new PVY detection tools clearly improve previously published PVY detection tests and offer new opportunities for PVY research programs.
authors.affiliation: INRA/ENSA, Unité Mixte de Recherche Biologie des Organismes et des Populations Appliquée à la Protection des Plantes (BiO3P), Domaine de la Motte B.P. 35327, F-35653 Le Rheu Cedex, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Balme-Sinibaldi
authors.forename: Michel
authors.initials: M
authors.lastname: Tribodet
authors.forename: Flora
authors.initials: F
authors.lastname: Croizat
authors.forename: Pierre
authors.initials: P
authors.lastname: Lefeuvre
authors.forename: Camille
authors.initials: C
authors.lastname: Kerlan
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Jacquot
mesh_terms: D017931:DNA Primers; D010935:Plant Diseases / Q000821:virology*; D020641:Polymorphism, Single Nucleotide; D017800:Potyvirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014026:Nicotiana / Q000821:virology; D010948:Viral Plaque Assay / Q000379:methods
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger