Real-Time PCR Assay Targeting the veA Gene for Quantification of Aspergillus carbonarius in Grapes.
J Food Prot
; 78(12): 2240-6, 2015 Dec.
Article
em En
| MEDLINE
| ID: mdl-26613920
ABSTRACT
In this work, a SYBR Green I real-time PCR method has been developed for the detection and quantification of Aspergillus carbonarius in grapes by targeting the veA gene with a primer pair (veAF4/veAR4) that specifically amplifies a 91-bp PCR product. The quantification of the fungal DNA was performed by generation of standard curves for two A. carbonarius strains, using spectrophotometrically measured DNA quantities (Log) with a linearity range from 50 to 5 × 10(-4) ng of DNA. A high positive correlation (R(2) > 0.99) between exponential increases of DNA and real-time PCR threshold cycles showed a high amplification efficiency for the assay (E values 100.06 and 101.51%, respectively). Quantification of the fungal genomic DNA in grape samples artificially inoculated with A. carbonarius conidia was successfully performed with a minimum threshold of 10(4) conidia per g of grape berry. The assay developed would allow reliable, specific, and efficient detection and quantification of A. carbonarius in grapes.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Aspergillus
/
DNA Fúngico
/
Vitis
/
Reação em Cadeia da Polimerase em Tempo Real
/
Frutas
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article