title: Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR.
abstract: Broad bean wilt virus 1 (BBWV-1) and BBWV-2 are the two most significant viruses in the genus Fabavirus, causing damage to many economically important agricultural crops worldwide. A quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) procedure using two TaqMan(®)MGB probes was developed for sensitive and specific detection and quantitation of BBWV-1 and BBWV-2. Primers and probes were designed from conserved sequence stretches to detect all isolates of each virus. Standard curves using RNA transcripts identical to both TaqMan(®)MGB probes enabled absolute quantitation, with a wide dynamic range and high sensitivity (10(3)-10(10) RNA molecules). RT-qPCR was assayed with genetically divergent BBWV-1 and BBWV-2 isolates from different plant hosts and countries, and was used to evaluate the temporal accumulation of BBWV-1 RNA in two plant hosts.
authors.affiliation: Instituto Valenciano de Investigaciones Agrarias, 46113 Moncada, Valencia, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Ferriol
authors.forename: S
authors.initials: S
authors.lastname: Ruiz-Ruiz
authors.forename: L
authors.initials: L
authors.lastname: Rubio
mesh_terms: D001483:Base Sequence; D027465:Chenopodium quinoa / Q000821:virology*; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D019181:Fabavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D015341:Nucleic Acid Probes / Q000235:genetics; D010935:Plant Diseases / Q000821:virology; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D031307:Vicia faba / Q000821:virology*
chemical_list: D017931:DNA Primers; D015341:Nucleic Acid Probes; D012367:RNA, Viral