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Differentiation between Aspergillus flavus and Aspergillus parasiticus from Pure Culture and Aflatoxin-Contaminated Grapes Using PCR-RFLP Analysis of aflR-aflJ Intergenic Spacer

El Khoury, André and Atoui, Ali and Rizk, Toufic and Lteif, Roger and Kallassy, Mireille and Lebrihi, Ahmed Differentiation between Aspergillus flavus and Aspergillus parasiticus from Pure Culture and Aflatoxin-Contaminated Grapes Using PCR-RFLP Analysis of aflR-aflJ Intergenic Spacer. (2011) Journal of Food Science , vol. 76 (n° 4). pp. M247-M253. ISSN 0022-1147

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Official URL: http://dx.doi.org/10.1111/J.1750-3841.2011.02153.X

Abstract

Aflatoxins (AFs) represent the most important single mycotoxin-related food safety problem in developed and developing countries as they have adverse effects on human and animal health. They are produced mainly by Aspergillus flavus and A. parasiticus. Both species have different aflatoxinogenic profile. In order to distinguish between A. flavus and A. parasiticus, gene-specific primers were designed to target the intergenic spacer (IGS) for the AF biosynthesis genes, aflJ and aflR. Polymerase chain reaction (PCR) products were subjected to restriction endonuclease analysis using BglII to look for restriction fragment length polymorphisms (RFLPs). Our result showed that both species displayed different PCR-based RFLP (PCR-RFLP) profile. PCR products from A. flavus cleaved into 3 fragments of 362, 210, and 102 bp. However, there is only one restriction site for this enzyme in the sequence of A. parasiticus that produced only 2 fragments of 363 and 311 bp. The method was successfully applied to contaminated grapes samples. This approach of differentiating these 2 species would be simpler, less costly, and quicker than conventional sequencing of PCR products and/or morphological identification.

Item Type:Article
Additional Information:Thanks to Wiley-Blackwell editor. The definitive version is available at http://onlinelibrary.wiley.com The original PDF of the article can be found at : http://onlinelibrary.wiley.com/doi/10.1111/j.1750-3841.2011.02153.x/abstract
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Centre National de la Recherche Scientifique - CNRS
Other partners > National Council For Scientific Research Lebanon - CNRS (LEBANON)
Université de Toulouse > Institut National Polytechnique de Toulouse - INPT
Université de Toulouse > Université Paul Sabatier-Toulouse III - UPS
Other partners > Université Saint-Joseph (LEBANON)
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Deposited By:Audrey LEFEVRE

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