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An innovative reverse line blot for simultaneous detection of animal species in food

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Animal products play a significant role in human consumption. Therefore, identifying the food fraud is of great importance. We describe a simple but an innovative method for the simultaneous detection… Click to show full abstract

Animal products play a significant role in human consumption. Therefore, identifying the food fraud is of great importance. We describe a simple but an innovative method for the simultaneous detection of meat sources from different animal species (cattle, buffalo, sheep, goat, horse, camel, pig, dog, cat, rabbit, chicken, wild duck and turkey) and human as a control DNA was extracted from samples prepared from human and above-mentioned animals and amplified using a common primer pair derived from a region of the mitochondrial DNA. The PCR products were subsequently hybridized with the species-specific DNA probes covalently bonded to a Biodyne C blotting membrane. The results showed that the PCR products generated from mammalian reacted as expected, to catch-all probes, mammalian catch-all probes and the corresponding species specific DNA probes but not with bird catch-all probes. The avian PCR product reacted also as expected, to catch-all probes, bird catch-all probes and the corresponding species-specific DNA probes, and not with the mammalian catch-all probes. These results showed that the used detection system could discriminate simultaneously, each animal species from the others. To determine the sensitivity of the presented hybridization method, the meat samples from 2 to 6 animals were mixed and the extracted DNA was analyzed. Interestingly, the species-specific reaction could be detected in all mixtures consisting of different species. Furthermore, it could be shown that the mixed meat DNA ratio 1/100 also could be detected simultaneously.

Keywords: dna; animal species; simultaneous detection; catch probes; food

Journal Title: European Food Research and Technology
Year Published: 2018

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