Aims: Development of a fast, sensitive and easy to handle method for the detection of Fusarium graminearum contamination in cereal samples by PCR. Methods and Results: DNA Detection Test StripsTM were used for PCR‐product detection and the method was compared to agarose gel electrophoresis. A minimum of 0·26 ng of purified target DNA was detectable with the Test StripTM Detection limit in less contaminated samples was slightly lower when gel electrophoresis was used for amplicon detection. In highly contaminated samples, detection limits of both methods were similar. Conclusions: Detection of PCR products was performed in 20 min without the need of special technical equipment or hazardous fluorescent DNA dyes. Significance and Impact of the Study: The new method described is useful for the screening of cereals in industrial quality control.

Several PCR methods have recently been developed for Fusarium analysis in pure cultures. To use these new techniques in mycological studies and in industrial quality control, a protocol was set up for the rapid preparation of fungal DNA from cereals. An ultrasonification probe (sonotrode) and a lysis buffer were used for mechanical lysis of mycelia from infected grains. Following ultrasonification, DNA was isolated using a commercially available kit. DNA preparation was completed within 5 min per sample. The method resulted in DNA of sufficient quality and quantity for diagnostic PCR. Group-and species-specific primers were used to detect DNA of Fusarium graminearum and F. culmorum in speciesspecific assays as well as trichothecene-producing Fusarium spp. in a group-specific system. A minimum of one F. graminearum -infected grain added to an uninfected 40 g wheat sample was detectable with a species-specific PCR. The PCR signals produced with primers specific for the tri5 gene of trichothecene-producing Fusarium spp. and with primers for the detection of F. graminearum (gaoA) were in accordance with corresponding concentrations of deoxynivalenol (DON) found in samples by HPLC analysis. The speed of the protocol developed may promote the use of PCR in routine applications in an agro-industrial context.

Trichothecenes pose a health risk to consumers and the presence of the producing fungi causes problems in industrial processing of cereals. A Triplex PCR was developed for screening of samples contaminated with toxigenic Fusarium species. The Triplex PCR could detect Fusarium species with high precision as compared to microbiological data of sample materials. There was a positive correlation between the occurrence of a PCR signal and the prevalence of the mycotoxin DON (Deoxynivalenol).