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

PloS One
Félix MoránAna B Ruiz-García

Abstract

Grapevine Pinot gris virus (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 108 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 Colomerus vitis. 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 poin...Continue Reading

References

Jan 16, 2007·Journal of Virological Methods·Fatima OsmanAdib Rowhani
Oct 18, 2013·Molecular Biology and Evolution·Koichiro TamuraSudhir Kumar
Jul 28, 2015·Scientific Reports·Piotr WysoczanskiMarkus Zweckstetter
Dec 17, 2015·Archives of Virology·Nadia BertazzonElisa Angelini
Jun 28, 2016·Archives of Virology·Valeria MalagniniValeria Gualandri
Jun 15, 2017·Virus Genes·Qi Wu, Nuredin Habili

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Datasets Mentioned

BETA
RQ125
RQ136
543887400

Methods Mentioned

BETA
PCR
in vitro transcription

Software Mentioned

LightCycler
Google Maps
Mega

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