title: Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR.
abstract: Tomato torrado virus (ToTV) causes serious damage to the tomato industry and significant economic losses. A quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR) method using primers and a specific TaqMan(®) MGB probe for ToTV was developed for sensitive detection and quantitation of different ToTV isolates. A standard curve using RNA transcripts enabled absolute quantitation, with a dynamic range from 10(4) to 10(10) ToTV RNA copies/ng of total RNA. The specificity of the RT-qPCR was tested with twenty-three ToTV isolates from tomato (Solanum lycopersicum L.), and black nightshade (Solanum nigrum L.) collected in Spain, Australia, Hungary and France, which covered the genetic variation range of this virus. This new RT-qPCR assay enables a reproducible, sensitive and specific detection and quantitation of ToTV, which can be a valuable tool in disease management programs and epidemiological studies.
authors.affiliation: Laboratorio de Protección Vegetal (LPV), Centro de Investigación Agropecuaria Central (CIAC), Instituto de Investigación Agropecuaria de Panamá (IDIAP), Ctra. Panamericana, Los Canelos, Santa María, Estafeta de Divisa, 0619 Herrera, Panama.
authors.forename: José Angel
authors.initials: JA
authors.lastname: Herrera-Vásquez
authors.affiliation: Instituto Valenciano de Investigaciones Agrarias (IVIA), 46113 Moncada, Valencia, Spain.
authors.forename: Luis
authors.initials: L
authors.lastname: Rubio
authors.affiliation: Grupo de Virología, Instituto Agroforestal Mediterráneo (IAM), Universidad Politécnica de Valencia (UPV), Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Alfaro-Fernández
authors.affiliation: Instituto Valenciano de Investigaciones Agrarias (IVIA), 46113 Moncada, Valencia, Spain.
authors.forename: Diana Elvira
authors.initials: DE
authors.lastname: Debreczeni
authors.affiliation: Grupo de Virología, Instituto Agroforestal Mediterráneo (IAM), Universidad Politécnica de Valencia (UPV), Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Font-San-Ambrosio
authors.affiliation: Department of Plant Pathology, University of California Davis, 95616 Davis, CA, United States.
authors.forename: Bryce W
authors.initials: BW
authors.lastname: Falk
authors.affiliation: Instituto Valenciano de Investigaciones Agrarias (IVIA), 46113 Moncada, Valencia, Spain; Department of Plant Pathology, University of California Davis, 95616 Davis, CA, United States. Electronic address: iferriolsafont@ucdavis.edu.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Ferriol
mesh_terms: D018551:Solanum lycopersicum / Q000821:virology; D010935:Plant Diseases / Q000821:virology; D010942:Plant Viruses / Q000302:isolation & purification*; D012328:RNA Viruses / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*
keywords: Detection; Diagnosis; Quantitative RT-PCR; Secoviridae; Torradovirus