Identification of Fusarium graminearum in cereal samples by DNA Detection Test StripsTM

Abstract

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.

The application of PCR in the detection of mycotoxigenic fungi in foods

“…The yield and purity of the extracted DNA could be improved by adapting the extraction procedure to the matrix of the DNA source and the results presented in the literature demonstrate the functionality of the developed PCR assays to detect mycotoxigenic moulds without an enrichment step by isolation of DNA directly from agricultural commodities, foods or animal feeds, such as maize (6), wheat (26), peanuts (8), and figs (16). In general, the sample material was frozen in liquid nitrogen, ground in a mortar and re-suspended in a lysis buffer.…”

“…Due to the rapidity and the possibility to handle multiple samples, the ultrasonic approach is also suitable for routine analysis. For purification of the extracted DNA the DNeasy Plant Mini kit from Qiagen has been widely used (8,20,26,38,45).…”

“…Furthermore, DNA dyes are hazardous for humans as they are potential mutagens. Therefore, novel system for a more rapid and convenient detection of PCR products have been developed (26,36,41). One of the systems is based on a PCR in which one of the primers is biotinylated and digoxigenin-11-dUTP is incorporated during elongation (41).…”

Microbial Diversity in Milled Rice as Revealed by Riosomal Intergenic Spacer Analysis

“…The yield and purity of the DNA isolated from foodstuffs are crucial determining the success of PCR-based microbial examinations, and a number of improvements to DNA extraction procedures have been reported 5,9,14,25) . However, most of these methods require a series of extraction and purification steps and an appreciable quantity of starting material.…”

“…As a result, they are not of practical use in the routine microbial examination of food commodities. For the purification of extracted DNA, commercial kits have also been widely used 9,16,25,37,46) . However, in our experience, these kits are not sufficiently robust to obtain high quality DNA from polysaccharide-rich materials, such as milled rice, and yield poor results during microbial community analyses (data not shown).…”