http://www.cnr.it/ontology/cnr/individuo/prodotto/ID56937

Development and validation of a multiplex RT-PCR method for the simultaneous detection of five grapevine viroids (Articolo in rivista)

Type

Prodotto della ricerca (Classe)
Articolo in rivista (Classe)

Label

Development and validation of a multiplex RT-PCR method for the simultaneous detection of five grapevine viroids (Articolo in rivista) (literal)

Anno

2012-01-01T00:00:00+01:00 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi

10.1016/j.jviromet.2011.09.022 (literal)

Alternative label

Hajizadeh M., Navarro B., Bashir N.S., Torchetti E.M., Di Serio F. (2012)
Development and validation of a multiplex RT-PCR method for the simultaneous detection of five grapevine viroids
in Journal of virological methods
(literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

Hajizadeh M., Navarro B., Bashir N.S., Torchetti E.M., Di Serio F. (literal)

Pagina inizio

62 (literal)

Pagina fine

69 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#url

http://www.sciencedirect.com/science/article/pii/S0166093411003971 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

179 (literal)

Rivista

Journal of virological methods (Rivista)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#note

IMPACT FACTOR RIVISTA: 2,139 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#pagineTotali

8 (literal)

Note

Scopu (literal)
ISI Web of Science (WOS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

Plant Protection Department, University of Tabriz, 29 Bahman, 51664, Tabriz, Iran (literal)

Titolo

Development and validation of a multiplex RT-PCR method for the simultaneous detection of five grapevine viroids (literal)

Abstract

Grapevine yellow speckle viroid 1 (GYSVd-1), Grapevine yellow speckle viroid 2 (GYSVd-2), Australian grapevine viroid (AGVd), Hop stunt viroid (HSVd) and Citrus exocortis viroid (CEVd) are the five viroids known to infect naturally grapevines. We developed a multiplex RT-PCR (mRT-PCR) method for the simultaneous detection of these five viroids and the amplification of the cDNA fragment of a host-derived mRNA (actin mRNA) as an internal positive control. Specific primers for each viroid targeted were designed by taking into account the sequence variability within and between the viroid species and tested in silico. The method was validated by testing 57 grapevine samples from Iran and showed reliability and high sensitivity. The RT-PCR-negative samples were further assayed by Northern-blot hybridization. For this, a method was developed for the simultaneous detection of three different grapevine viroids on a single hybridization membrane. In this survey, HSVd, AGVd, GYSVd-1, and GYSVd-2 were detected in 100, 95, 93, and 65% of the samples tested, respectively, confirming the wide distribution of these viroids in Iran. CEVd was not detected in any of the samples collected. Based on these results, HSVd is proposed as a positive internal control for mRT-PCR in the areas where this viroid is widespread, so as to reduce the time and costs of DNase treatment, which is required when a host-derived internal control is used. The mRT-PCR method has the potential to be used routinely for large-scale surveys and certification programs. (literal)

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