21034774
not annotated - annotated - LINNAEUS only
Detection of Tomato black ring virus by real-time one-step RT-PCR.
A TaqMan-based real-time one-step RT-PCR assay was developed for the rapid detection of Tomato black ring virus (TBRV), a significant plant pathogen which infects a wide range of economically important crops. Primers and a probe were designed against existing genomic sequences to amplify a 72 bp fragment from RNA-2. The assay amplified all isolates of TBRV tested, but no amplification was observed from the RNA of other nepovirus species or healthy host plants. The detection limit of the assay was estimated to be around nine copies of the TBRV target region in total RNA. A comparison with conventional RT-PCR and ELISA, indicated that ELISA, the current standard test method, lacked specificity and reacted to all nepovirus species tested, while conventional RT-PCR was approximately ten-fold less sensitive than the real-time RT-PCR assay. Finally, the real-time RT-PCR assay was tested using five different RT-PCR reagent kits and was found to be robust and reliable, with no significant differences in sensitivity being found. The development of this rapid assay should aid in quarantine and post-border surveys for regulatory agencies.
Ann file
T1 Species 13 36 Tomato black ring virus
N1 Reference T1 Taxonomy:12275
T2 Species 156 179 Tomato black ring virus
N2 Reference T2 Taxonomy:12275
T3 Species 181 185 TBRV
N3 Reference T3 Taxonomy:12275
T4 Species 422 426 TBRV
N4 Reference T4 Taxonomy:12275
T5 Species 612 616 TBRV
N5 Reference T5 Taxonomy:12275