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A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites

Felix Moran; Antonio Olmos; Leonidas Lotos; Lukas Predajňa; Nikolaos Katis; Miroslav Glasa; Varvara Maliogka; Ana B. Ruiz-Garcia


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    "doi": "10.1371/journal.pone.0197237", 
    "description": "<p><em>Grapevine Pinot gris virus</em> (GPGV) is a widely distributed grapevine pathogen that has been associated to the grapevine leaf mottling and deformation disease. With the aim of better understanding the disease epidemiology and providing efficient control strategies a specific and quantitative duplex TaqMan real-time RT-PCR assay has been developed. This method has allowed reliable quantitation of the GPGV titer ranging from 30 up to 3 x 10<sup>8</sup> transcript copies, with a detection limit of 70 viral copies in plant material. The assay targets a grapevine internal control that reduces the occurrence of false negative results, thus increasing the diagnostic sensitivity of the technique. Viral isolates both associated and non-associated to symptoms from Greece, Slovakia and Spain have been successfully detected. The method has also been applied to the absolute quantitation of GPGV in its putative transmission vector <em>Colomerus vitis</em>. Moreover, the viral titer present in single mites has been determined. In addition, in the current study a new polymorphism in the GPGV genome responsible for a shorter movement protein has been found. A phylogenetic study based on this genomic region has shown a high variability among Spanish isolates and points to a different evolutionary origin of this new polymorphism. The methodology here developed opens new possibilities for basic and epidemiological studies as well as for the establishment of efficient control strategies.</p>", 
    "language": "eng", 
    "title": "A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites", 
    "license": {
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      "pages": "e0197237", 
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    "keywords": [
      "GPGV, Grapevine, Real-time RT-PCR, Quantification"
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    "publication_date": "2018-05-15", 
    "creators": [
      {
        "affiliation": "Centro de Proteccion Vegetal y Biotecnologia, Instituto Valenciano de Investigaciones Agrarias (IVIA), Moncada, Valencia, Spain", 
        "name": "Felix Moran"
      }, 
      {
        "affiliation": "Centro de Proteccion Vegetal y Biotecnologia, Instituto Valenciano de Investigaciones Agrarias (IVIA), Moncada, Valencia, Spain", 
        "name": "Antonio Olmos"
      }, 
      {
        "affiliation": "Aristotle University of Thessaloniki, Faculty of Agriculture, Forestry and Natural Environment, School of Agriculture, Plant Pathology Laboratory, Thessaloniki, Greece", 
        "name": "Leonidas Lotos"
      }, 
      {
        "affiliation": "Institute of Virology, Biomedical Research Centre, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, Slovakia", 
        "name": "Lukas Predaj\u0148a"
      }, 
      {
        "affiliation": "Aristotle University of Thessaloniki, Faculty of Agriculture, Forestry and Natural Environment, School of Agriculture, Plant Pathology Laboratory, Thessaloniki, Greece", 
        "name": "Nikolaos Katis"
      }, 
      {
        "affiliation": "Institute of Virology, Biomedical Research Centre, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, Slovakia", 
        "name": "Miroslav Glasa"
      }, 
      {
        "affiliation": "Aristotle University of Thessaloniki, Faculty of Agriculture, Forestry and Natural Environment, School of Agriculture, Plant Pathology Laboratory, Thessaloniki, Greece", 
        "name": "Varvara Maliogka"
      }, 
      {
        "affiliation": "Centro de Proteccion Vegetal y Biotecnologia, Instituto Valenciano de Investigaciones Agrarias (IVIA), Moncada, Valencia, Spain", 
        "name": "Ana B. Ruiz-Garcia"
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