Highly sensitive PCR-based detection method specific to Aspergillus flavus in wheat flour
Résumé
Aspergillus flavus is an important fungal species which frequently occurs in food commodities producing a wide array of toxins, being aflatoxins the most relevant in Food Safety. In this work, a PCR-based protocol is described specific to this specie which allowed discrimination from other closely related species having different profiles of secondary metabolites from the Aspergillus Section Flavi, particularly A. parasiticus. The specific primers were designed on the multicopy internal transcribed region of the rDNA unit (ITS1-5.8S-ITS2 rDNA) and were tested in a wide sample of related species and other fungal species commonly found in commodities. The PCR assay was coupled with a fungal enrichment and a DNA extraction method for wheat flour to enhance the sensitivity of the diagnostic protocol. The results indicated that the critical PCR amplification product was clearly observed for wheat flour contaminated by 102 spores after 16 h of incubation.
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