Abstract:
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.
