Journal article Open Access

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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{
  "note": "This publication reflects only the authors' view. The Agency is not responsible for any use that may be made of the information it contains.", 
  "DOI": "10.1371/journal.pone.0197237", 
  "container_title": "PLoS ONE", 
  "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", 
  "issued": {
    "date-parts": [
      [
        2018, 
        5, 
        15
      ]
    ]
  }, 
  "abstract": "<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>", 
  "author": [
    {
      "family": "Felix Moran"
    }, 
    {
      "family": "Antonio Olmos"
    }, 
    {
      "family": "Leonidas Lotos"
    }, 
    {
      "family": "Lukas Predaj\u0148a"
    }, 
    {
      "family": "Nikolaos Katis"
    }, 
    {
      "family": "Miroslav Glasa"
    }, 
    {
      "family": "Varvara Maliogka"
    }, 
    {
      "family": "Ana B. Ruiz-Garcia"
    }
  ], 
  "page": "e0197237", 
  "volume": "13", 
  "type": "article-journal", 
  "issue": "5", 
  "id": "1248646"
}
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