Validation of a Quick PCR Method Suitable for Direct Sequencing: Identification of Fusarium Fungal Species and Chemotypes for Preventive Approaches in Food Safety

Marine Pallez, Matias Pasquali*, Torsten Bohn, Lucien Hoffmann and Marco Beyer

Public Research Centre Gabriel Lippmann, Environment and Agro-Biotechnologies Department, 41, rue du Brill, LU-4422 Belvaux, Luxembourg

Article history:
Received April 26, 2013
Accepted February 19, 2014

Key words:
Fusarium culmorum, Fusarium graminearum, microwave DNA extraction, EF1-α

Summary:
Species determination by sequencing and PCR genetic chemotyping, used to determine the toxigenic potential of Fusarium strains, is fundamental for developing preventive strategies in food safety. Here we propose and statistically validate a quick protocol for standardizing the procedure of species determination by sequencing of the elongation factor 1-α and multiplex genetic chemotyping using the Tri12 gene, based on fungal growth on Miracloth tissue coupled with microwave extraction. The test was validated on 75 Fusarium culmorum and Fusarium graminearum strains.

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