Development of Rt-PCR Assays Using Fluorogenic 3' Minor Groove Binder Dna Probes for Detection of Fruit Tree Viruses

Marbot, Sophie; Kummert, J.; Vendrame, Marina; Dutrecq, O.; Lepoivre, Philippe; Salmon, Michel

doi:10.17660/actahortic.2004.657.89

Cited by 1 publication

(3 citation statements)

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“…size of the probe (Marbot et al, 2004;Osman et al, 2007). Virus detection through real-time RT-PCR (TaqMan) has been confirmed to be highly sensitive, specific and robust, being able to detect different viruses from collected samples in Brazilian vineyards.…”

Section: Ampelovirusmentioning

confidence: 99%

“…This is particularly true when mutation or nucleotide substitution occurs within the probe pairing site, resulting in false-negative (James et al, 2006). Marbot et al (2004) demonstrated that a mismatch of a single nucleotide between probe and the target viral sequence may prevent the fluorogenic reaction from proceeding. Sequencing of a large number of isolates may allow the design of probes in more conserved regions.…”

Section: Ampelovirusmentioning

confidence: 99%

“…Molecular techniques such as reverse transcription polymerase chain reaction (RT-PCR) can surpass ELISA in sensitivity for detection of some viruses in dormant grapevines. However, this method requires gel electrophoresis analysis for its interpretation and this post-detection step has hampered the routine use of the RT-PCR technique (Marbot et al, 2004).…”

mentioning

confidence: 99%

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

Dubiela¹,

Fajardo²,

Souto³

et al. 2013

Trop. plant pathol.

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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.

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Section: Ampelovirusmentioning

confidence: 99%

Section: Ampelovirusmentioning

confidence: 99%