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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
Author(s) -
El Khoury André,
Atoui Ali,
Rizk Toufic,
Lteif Roger,
Kallassy Mireille,
Lebrihi Ahmed
Publication year - 2011
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/j.1750-3841.2011.02153.x
Subject(s) - restriction fragment length polymorphism , aspergillus flavus , biology , aspergillus parasiticus , restriction enzyme , intergenic region , aflatoxin , polymerase chain reaction , genetics , microbiology and biotechnology , gene , genome , food science
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.