First molecular characterization of grapevine virus B (GVB) in Portuguese grapevine cultivars and improvement of the RT-PCR detection assay

This work describes the first molecular characterization of grapevine virus B (GVB) in Portuguese grapevine cultivars. During a routine screening of 44 accessions in the National Collection of Grapevine Varieties (CAN PRT051), 17 were found infected with GVB in DAS-ELISA assays with commercial antib...

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Bibliographic Details
Published in:Archives of virology 2016-12, Vol.161 (12), p.3535-3540
Main Authors: Fonseca, Filomena, Duarte, Vilma, Teixeira Santos, Margarida, Brazão, João, Eiras-Dias, Eduardo
Format: Article
Language:English
Subjects:
Antibodies
Biomedical and Life Sciences
Biomedicine
Brief Report
Cloning
Cultivars
DNA Primers - genetics
Flexiviridae - classification
Flexiviridae - genetics
Flexiviridae - isolation & purification
Genetic Variation
Grapevine virus B
Infectious Diseases
Medical Microbiology
Open Reading Frames
Phylogenetics
Phylogeny
Plant Diseases - virology
Portugal
Proteins
Reverse Transcriptase Polymerase Chain Reaction - methods
Sequence Analysis, DNA
Serology
Virology
Virology - methods
Vitaceae
Vitis - virology
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Summary:This work describes the first molecular characterization of grapevine virus B (GVB) in Portuguese grapevine cultivars. During a routine screening of 44 accessions in the National Collection of Grapevine Varieties (CAN PRT051), 17 were found infected with GVB in DAS-ELISA assays with commercial antibodies. However, only six of the corresponding isolates were successfully amplified using primer pairs described in the literature. The sequence variants (ORF4-3’UTR, 1147 nt) retrieved from these isolates segregated into two phylogenetic groups, which included sequences from complete genomes available in GenBank. The highly discrepant results obtained using serological and RT-PCR-based diagnostic tools led to the design of a primer pair for detection of GVB, which allowed the amplification of a 606-bp GVB-specific fragment from all DAS-ELISA-positive isolates and also revealed the existence of false negatives in the serological testing.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-016-3024-3