title: Detection and discrimination of European isolates of Soil-borne wheat mosaic virus using immunocapture real-time reverse transcription-polymerase chain reaction.
abstract: Soil-borne wheat mosaic virus (SBWMV) is a viral pathogen of cereal crops. Although first detected in the United States, it has since been reported in Germany and Poland. To date, two SBWMV strains (Nebraska-like and New-York-like types) have been detected. While the nucleotide sequences of the coat protein (CP) genes from the two strains show only 89% similarity, the deduced amino acid sequences of this region are identical. Therefore, the isolates cannot be differentiated serologically using the double antibody sandwich enzyme-linked immunosorbent assay. An immunocapture real-time reverse transcription-polymerase chain reaction (IC-real-time-RT-PCR) assay was developed for detection and discrimination of SBWMV strains based on the amplification of short CP fragments (∼250bp). The results showed distinct melting curve profiles related to each strain. The primer pairs gave similar melting points, with a Tm of 82.1°C for Nebraska-like and 83.5°C for New-York-like SBWMV strains. This work confirms that IC-real-time-RT-PCR is a valuable technique for rapid detection and discrimination of SBWMV strains.
authors.affiliation: Institute of Plant Protection, National Research Institute, Department of Virology and Bacteriology, Władysława Węgorka 20, 60-318 Poznań, Poland. Electronic address: k.trzmiel@iorpib.poznan.pl.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Trzmiel
authors.affiliation: Institute of Plant Protection, National Research Institute, Department of Virology and Bacteriology, Władysława Węgorka 20, 60-318 Poznań, Poland.
authors.forename: Marzena
authors.initials: M
authors.lastname: Lewandowska
mesh_terms: D036022:Capsid Proteins / Q000235:genetics; D005858:Germany; D007118:Immunoassay / Q000379:methods*; D010935:Plant Diseases / Q000821:virology; D010942:Plant Viruses / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011044:Poland; D012328:RNA Viruses / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012988:Soil Microbiology; D044366:Transition Temperature; D014908:Triticum / Q000821:virology; D014481:United States
chemical_list: D036022:Capsid Proteins; D012367:RNA, Viral
keywords: IC-real-time-RT-PCR; Soil-borne wheat mosaic virus; Strain determination