Improvement of Potato virus Y (PVY) detection and quantitation using PVY(N)- and PVY(O)-specific real-time RT-PCR assays.
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.
Balme-Sinibaldi
Valérie
V
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.
Tribodet
Michel
M
Croizat
Flora
F
Lefeuvre
Pierre
P
Kerlan
Camille
C
Jacquot
Emmanuel
E
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
D017931:DNA Primers; D012333:RNA, Messenger