Developing a PCR- based method for identification of Aspergillus flavus

Abstract

Owning the ability to grow in a wide range of humidity and temperature, Aspergillus flavus is most widely reported as foodborne fungus. It is frequently found in food; producing aflatoxin (e.g. AFB1, AFB2), A. flavus contaminations threat the food safety. A. flavus, together with Asperillus parasiticus, are also the major pathogenic Aspergillus species. The present study developed a fast PCR-based method for identification of A. flavus with high specificity and sensitivity. The specific primers were designed relying on the comparison of aflatoxin biosynthesis gene clusters of various Aspergillus species; and were tested on standard reference strains. Four DNA isolation protocols for molds were also tested. The limits of detection (LOD) were established with 0.008ng/µl for individual DNA concentration and 0.016ng/µl for mixture DNA concentration.

Key words: Aspergillus flavus, aflatoxin, PCR