Real-time RT-PCR (TaqMan®) assays for the detection of Grapevine Leafroll associated viruses 1–5 and 9

Osman, Fatima; Leutenegger, Christian M.; Golino, Deborah A.; Rowhani, Adib

doi:10.1016/j.jviromet.2006.11.035

Cited by 153 publications

(97 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The total RNA quality was monitored by inclusion of primers and probe for 18S rRNA (Osman et al, 2007). In all performed analyses, duplicates of RNAse free water, healthy grapevines and positive controls were included.…”

Section: Ampelovirusmentioning

confidence: 99%

“…Primer and probe sets are described in Table 1. For GLRaV-1 three sets of primers and probes were tested: Osman et al (2007); mixture of Osman et al (2007) and Klaassen et al (2011), a wide range reagent targeted to the same viral region; and Pacifico et al (2011) ( Table 1). All probes were labeled with fluorophores 6-FAM or VIC at their 5' end.…”

Section: Ampelovirusmentioning

confidence: 99%

“…Real-time RT-PCR has been used for detection of viruses that infect different host plants (James et al, 2006), including grapevines (Osman et al, 2007;Pacifico et al, 2011), having higher sensitivity than conventional RT-PCR, overcoming limitations of conventional virus detection systems affected by the erratic distribution of viruses in the host, variations in virus titre throughout the season and type of sampling tissue, as observed in grapevines. Additionally TaqMan probes can be labeled with different fluorophores, excited at different wavelengths, allowing the simultaneous detection of more than one virus species in a single reaction.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Simultaneous detection of Brazilian isolates of grapevine viruses by TaqMan real-time RT-PCR

Dubiela¹,

Fajardo²,

Souto³

et al. 2013

Trop. plant pathol.

View full text Add to dashboard Cite

The aim of this work was to evaluate the efficiency of real-time RT-PCR for detection of different isolates of ten important virus species that infect grapevines in Brazil: Grapevine leafroll-associated virus (GLRaV-1, -2, -3 and -5), Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine virus D (GVD), Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine fleck virus (GFkV) and Grapevine fanleaf virus (GFLV). The reactions consisted of individual (simplex) and simultaneous (duplex) virus detections. Thirty six grapevine accessions, regenerated after thermotherapy and tissue culture treatments, have been analysed. All the above-mentioned viruses were sensitively detected in simplex reactions in samples infected with different virus isolates. Specifically to GLRaV-1 it was necessary to mix reagents refered by different sources to achieve the amplification. GVA, GRSPaV, GLRaV-2 and GLRaV-3 combined with GVB, GFLV, GFkV, GVD and GLRaV-5 were accurately detected in duplex trials. It was shown, that real-time RT-PCR (TaqMan) is able to efficiently detect different local virus species and isolates.

show abstract

Section: Ampelovirusmentioning

confidence: 99%

Section: Ampelovirusmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Simultaneous detection of Brazilian isolates of grapevine viruses by TaqMan real-time RT-PCR

Dubiela¹,

Fajardo²,

Souto³

et al. 2013

Trop. plant pathol.

View full text Add to dashboard Cite

show abstract

“…All plant samples were extracted and tested for GLRaV −1, −2, −3 and -4LV as described in Osman et al (2007). Field-collected samples were also tested for GLRaV-3 using the CP primer set (Sharma et al 2011), which can detect some variants of GLRaV-3 not detected by the primers in Osman et al (2007).…”

mentioning

confidence: 99%

“…Field-collected samples were also tested for GLRaV-3 using the CP primer set (Sharma et al 2011), which can detect some variants of GLRaV-3 not detected by the primers in Osman et al (2007). Controls for GLRaV detection in plant material were roots and petioles of known GLRaV-positive or negative plants, and an extraction buffer blank with no plant material.…”

mentioning

confidence: 99%