Comparison of low-density arrays, RT-PCR and real-time TaqMan® RT-PCR in detection of grapevine viruses

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

doi:10.1016/j.jviromet.2008.01.012

Cited by 99 publications

(45 citation statements)

References 20 publications

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“…This suggests the poor sensitivity afforded by ELISA tests and the necessity to improve the reliability of detection methods for sanitary programs. The detection of viruses in grapevine is difficult because there are seasonal variations in the concentration of viruses and ELISA tests are not sensitive enough and reliable in different seasons, which agreed with reports by other authors (Osman et al 2008). The third consideration is that these high prevalences of viruses cannot be explained only by vector transmission and suggest that initial infected plant material and uncontrolled traffic of propagation material, were key factors for these high levels of infection.…”

supporting

confidence: 70%

“…On the other hand the design of specific primers and one TaqMan probe based on all isolates included in GenBank database allowed the amplification of a 146 bp product and the specific detection of all tested GFkV isolates. In the case of GFLV, GLRaV-1 and GLRaV-3 viruses, specific primers and TaqMan probes previously described were used (Osman et al 2008). Table 1 summarised specific primers and TaqMan probes used in this study.…”

mentioning

confidence: 99%

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High prevalence of viruses in table grape from Spain detected by real-time RT-PCR

Bertolini

García²,

Yuste

et al. 2010

Eur J Plant Pathol

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Table grapes from one of the most important growing area in Spain (Vinalopó, Alicante) protected by the Designation of Origin "Vinalopó bagged table grape", were surveyed and analysed to determine the prevalence of the five viruses included in the Spanish certification program: Arabis mosaic virus (ArMV), Grapevine fanleaf virus (GFLV), Grapevine fleck virus (GFkV), Grapevine leafroll associated virus-1 (GLRaV-1) and Grapevine leafroll associated virus-3 (GLRaV-3). Ninety five sampling points were selected and the position of grapevine plants georeferenced. Samples were collected in two different vegetative periods and analyses were performed by ELISA and real-time RT-PCR. Purified RNA and immobilized viral targets from plant extracts on nylon membranes were used in parallel assays as templates for PCR assays. In order to analyse these five viral species by real-time RT-PCR, new specific primers and TaqMan probes were designed for detection of ArMV and GFkV. Real time RT-PCR from purified RNA was more sensitive than spot version and ELISA tests. The most prevalent virus was GFLV (95.8%) followed by GLRaV-3 (94.7%), GLRaV-1 (66.3%) and GFkV (65.3%). ArMV was not detected in any sample. The high level of viral infections and the presence of mixed infections suggest that initial infected plant material and uncontrolled traffic of propagation material have played an important role in the spread of viruses.

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supporting

confidence: 70%

mentioning

confidence: 99%

High prevalence of viruses in table grape from Spain detected by real-time RT-PCR

Bertolini

García²,

Yuste

et al. 2010

Eur J Plant Pathol

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“…PCR, using detection chemistries such as TaqMan, is increasingly being used for the detection and quantification of GLRaVs in plant tissue and insect vectors compared to conventional RT-PCR assays (127). Low-density PCR arrays (LDA) using the real-time TaqMan PCR primers/probes complexes in 384-well plates were developed for the simultaneous detection and quantification of different GLRaVs (128). Overall, the TaqMan-based real time RT-PCR and LDA detection methods are rapid and quantitative, and can provide the required robustness in processing a large number of samples for the detection of GLRaVs (129).…”

Section: Diagnosismentioning

confidence: 99%

Grapevine Leafroll: A Complex Viral Disease Affecting a High-Value Fruit Crop

et al. 2014

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Grapevine (Vitis spp.) is one of the most widely grown fruit crops in the world. It is a deciduous woody perennial vine for which the cultivation of domesticated species began approximately 6,000 to 8,000 years ago in the Near East. Grapevines are broadly classified into red- and white-berried cultivars based on their fruit skin color, although yellow, pink, crimson, dark blue, and black-berried cultivars also exist. Grapevines can be subject to attacks by many different pests and pathogens, including graft-transmissible agents such as viruses, viroids, and phytoplasmas. Among the virus and virus-like diseases, grapevine leafroll disease (GLD) is by far the most widespread and economically damaging viral disease of grapevines in many regions around the world. The global expansion of the grape and wine industry has seen a parallel increase in the incidence and economic impact of GLD. Despite the fact that GLD was recognized as a potential threat to grape production for several decades, our knowledge of the nature of the disease is still quite limited due to a variety of challenges related to the complexity of this virus disease, the association of several distinct GLD-associated viruses, and contrasting symptoms in red- and white-berried cultivars. In view of the growing significance of GLD to wine grape production worldwide, this feature article provides an overview of the state of knowledge on the biology and epidemiology of the disease and describes management strategies currently deployed in vineyards.

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“…Several different methods such as biological indexing (Ahmed et al, 2004;Salem et al, 2007), serological assays (ELISA) (Lunden et al, 2010) and/or molecular assays (RT-PCR, multiplex PCR and real time PCR) (Gambino and Gribaudo, 2006;Osman et al, 2008;Wang et al, 2011) were previously used for detecting grape vine viruses. Our study for virus identification was, however, based solely on their detection by RT-PCR using virus-specific primers.…”

Section: Discussionmentioning

confidence: 99%

Detection and molecular characterization of Egyptian isolates of grapevine viruses

Fattouh¹,

Ratti²,

ElAhwany³

et al. 2014

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Comparison of low-density arrays, RT-PCR and real-time TaqMan® RT-PCR in detection of grapevine viruses

Cited by 99 publications

References 20 publications

High prevalence of viruses in table grape from Spain detected by real-time RT-PCR

High prevalence of viruses in table grape from Spain detected by real-time RT-PCR

Grapevine Leafroll: A Complex Viral Disease Affecting a High-Value Fruit Crop

Detection and molecular characterization of Egyptian isolates of grapevine viruses

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