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Early detection of ochratoxigenic fungi in wine grapes and of ochratoxin A in wine

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Abstract

The main objective of this work was to develop a protocol for the detection and quantification of ochratoxin A (OTA) to be used directly in food commodities, where sensitivity and robustness are critical factors. In this study, the contamination of wine grapes (var. Merlot) by OTA producer Aspergillus carbonarius was detected by PCR on asymptomatic grape berries. Four varieties of Vitis vinifera harvested directly from the field were used in the PCR-based assay. In the experiments, A. carbonarius was used to develop the PCR method, and Aspergillus niger strain 7096 and other species isolated from grapes were also used to test the specificity of the method presented below. The multiplex PCR reaction was tested on fungal DNA by amplifying two target genes in order to differentiate with an higher degree of effectiveness potentially ochratoxigenic black Aspergilli from other fungal contaminants. These PCR-based assays showed high sensitivity and effectiveness in detecting A. carbonarius from grape DNA. The same grape berries in which A. carbonarius DNA was revealed at an early timepoint showed the visible presence of black rot after appropriate incubation, confirming the results obtained by the molecular methods developed here. Furthermore, OTA detection on Merlot wine was performed with a novel device based on the use of an amorphous silica photosensor. This technique, using very simplified and rapid extraction procedures, allowed the detection and quantification of OTA from wine contaminated at 2 ppb. These methods allow rapid and early detection of the presence of the pathogen and of the toxin within a working day.

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Acknowledgements

This work was supported by National Project Me.Di.T.A. (MIUR code n. 9038) and by EU project no. 232245 “Novel photosensor-based device for rapid and quantitative ochratoxin A determination in wine, beer and feed”.

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Authors and Affiliations

  1. ENEA, CR-CASACCIA, Via Anguillarese 301, 00123, Rome, Italy

    Patrizia De Rossi

  2. Istituto di Chimica Biomolecolare-CNR, P.le Aldo Moro 5, 00185, Rome, Italy

    Alessandra Ricelli

  3. Dip. Biologia Vegetale, Università degli Studi “Sapienza”, Largo Cristina di Svezia 24, 00165, Rome, Italy

    Massimo Reverberi, Anna Adele Fabbri & Corrado Fanelli

  4. Dip. Ingegneria Elettronica, Università degli Studi “Sapienza”, Via Eudossiana, 18, 00184, Roma, Italy

    Domenico Caputo, Gianpiero De Cesare & Riccardo Scipinotti

Authors
  1. Patrizia De Rossi
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  2. Massimo Reverberi
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  3. Alessandra Ricelli
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  4. Anna Adele Fabbri
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  5. Domenico Caputo
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  6. Gianpiero De Cesare
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  7. Riccardo Scipinotti
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  8. Corrado Fanelli
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Corresponding author

Correspondence to Patrizia De Rossi.

Additional information

This paper is part of the special issue “Wine microbiology and safety: from the vineyard to the bottle (Microsafetywine)”, 19–20 November 2009, Martina Franca (Italy).

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De Rossi, P., Reverberi, M., Ricelli, A. et al. Early detection of ochratoxigenic fungi in wine grapes and of ochratoxin A in wine. Ann Microbiol 61, 11–15 (2011). https://doi.org/10.1007/s13213-010-0107-3

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  • DOI: https://doi.org/10.1007/s13213-010-0107-3

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