Real-time RT-PCR for detection of Raspberry bushy dwarf virus, Raspberry leaf mottle virus and characterizing synergistic interactions in mixed infections Public Deposited

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  • Two TaqMan-based real-time One-Step RT-PCR assays were developed for the rapid and efficient detection of Raspberry bushy dwarf virus (RBDV) and Raspberry leaf mottle virus (RLMV), two of the most common raspberry viruses in North America and Europe. The primers and probes were designed from conserved fragments of the polymerase region of each virus and were effective for the detection of different isolates tested in this study. The RBDV assay amplified a 94 bp amplicon and was able to detect as few as 30 viral copies. Whereas the RLMV assay amplified a 180 bp amplicon and detected as few as 300 viral copies from plant and aphid RNA extracts. Both assays were significantly more sensitive than their corresponding conventional RT-PCR methods. The sensitivity of the RLMV assay was also tested on single aphids after a fixed acquisition access period (AAP). In addition, the assays revealed a novel synergistic interaction between the two viruses, where the concentration of RBDV was enhanced ∼400-fold when it occurred in combination with RLMV compared to its concentration in single infections. The significance of this finding and the importance of the development of real-time RT-PCR assays for the detection of RBDV and RLMV are discussed.
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  • Quito-Avila, D. F., & Martin, R. R. (2012, January). Real-time RT-PCR for detection of Raspberry bushy dwarf virus, Raspberry leaf mottle virus and characterizing synergistic interactions in mixed infections. Journal of Virological Methods, 179(1), 38-44. doi:10.1016/j.jviromet.2011.09.016
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  • description.provenance : Approved for entry into archive by Deanne Bruner(deanne.bruner@oregonstate.edu) on 2012-05-30T23:14:28Z (GMT) No. of bitstreams: 1 Quito-AvilaDiegoBotanyPlantPathologyRealTimeDetection.pdf: 962286 bytes, checksum: 97b248634939f4d102bb08f6b71227d6 (MD5)
  • description.provenance : Submitted by Deanne Bruner (deanne.bruner@oregonstate.edu) on 2012-05-30T19:25:51Z No. of bitstreams: 1 Quito-AvilaDiegoBotanyPlantPathologyRealTimeDetection.pdf: 962286 bytes, checksum: 97b248634939f4d102bb08f6b71227d6 (MD5)
  • description.provenance : Made available in DSpace on 2012-05-30T23:14:28Z (GMT). No. of bitstreams: 1 Quito-AvilaDiegoBotanyPlantPathologyRealTimeDetection.pdf: 962286 bytes, checksum: 97b248634939f4d102bb08f6b71227d6 (MD5) Previous issue date: 2012-01

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