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Naue J.,Albert Ludwigs University of Freiburg | Lutz-Bonengel S.,Albert Ludwigs University of Freiburg | Pietsch K.,State Institute for Chemical and Veterinary Analysis of Food Freiburg | Sanger T.,Albert Ludwigs University of Freiburg | And 2 more authors.
International Journal of Legal Medicine | Year: 2012

The authors report on a young boy who was bitten into his face by an unknown animal while being asleep in a tent. Given the bite marks and the location of the scene, members of the mustelidae and canidae families were the first "suspects." Deoxyribunucleic acid (DNA) recovered from the tent's wall was analyzed with regard to parts of the mitochondrial 12S ribosomal ribunucleic acid (12S rRNA) and cytochrome b (cytb) genes as well as nuclear short tandem repeats (STRs). Since Sanger sequencing revealed a mixed sequence with a strong human component overlying the nonhuman contributor, an animal screening using a duplex real-time polymerase chain reaction (PCR) with an intercalating dye and melt curve analysis was employed. The results were later confirmed by cloning. The applied commercial canine STR kit verified the animal family (canidae) but did not help in discriminating the species due to cross-species amplification. In the presented case, the real-time PCR assay offered the cheapest and fastest method for animal family determination, which then allowed for an appropriate and sample-saving strategy to characterize the causative animal species. © Springer-Verlag 2012.

Waiblinger H.-U.,State Institute for Chemical and Veterinary Analysis of Food Freiburg | Grohmann L.,Federal Office of Consumer Protection and Food Safety | Mankertz J.,Federal Office of Consumer Protection and Food Safety | Engelbert D.,Federal Office of Consumer Protection and Food Safety | Pietsch K.,State Institute for Chemical and Veterinary Analysis of Food Freiburg
Analytical and Bioanalytical Chemistry | Year: 2010

In routine analysis, screening methods based on real-time PCR are most commonly used for the detection of genetically modified (GM) plant material in food and feed. In this paper, it is shown that the combination of five DNA target sequences can be used as a universal screening approach for at least 81 GM plant events authorised or unauthorised for placing on the market and described in publicly available databases. Except for maize event LY038, soybean events DP-305423 and BPS-CV127-9 and cotton event 281-24-236 × 3006-210-23, at least one of the five genetic elements has been inserted in these GM plants and is targeted by this screening approach. For the detection of these sequences, fully validated real-time PCR methods have been selected. A screening table is presented that describes the presence or absence of the target sequences for most of the listed GM plants. These data have been verified either theoretically according to available databases or experimentally using available reference materials. The screening table will be updated regularly by a network of German enforcement laboratories. © 2009 Springer-Verlag.

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