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Home » Archive » 2014

TDK conference 2014

Cow’s milk detection from wine using molecular methods
Csősz György - year 4
SZIU Faculty of Veterinary Science, Institute for Animal Breeding, Nutrition and Laboratory Animal Science, Department of Animal Breeding and Genetics
Supervisor: Dr. Ákos Maróti-Agóts

Abstract:

During milk secretion that located in the mammary gland, mitochondrions could appear in milk, as a result of many physiological and pathological mechanisms. Physiologically, by natural types of milk secretion, pathologically by the increased amount of somatic cells. The mitochondrions contain mitochondrial DNA , due to their origin and function. Cow's milk which contains the mitochondrions of cattle appears in wine for various reasons. Milk could be used for coagulation of floating, or dissolved particles, and for refinement of acids, though milk could get in wine as contamination. Milk contamination can appear as a result of special wine falsification method, when poor quality wine from abroad comes to our homeland, in tank trucks, that were used for milk logistics in reverse direction before. Fakers mix this quantity of wine with Hungarian wine, having the same poor quality as the ilegally imported one. Detection of milk from wine made by this falsification method propounds the question of regular and thorough cleaning of tank trucks.

In our paper, we were looking for the answer, what level of sensitivity could we detect cow’s milk from wine, using simple molecular methods (DNA cleaning, PCR, QPCR). In cooperation with the National Food Chain Safety Office’s Direction of Wine and Alcoholic Drinks, we examined clear wines from noted Hungarian producers, and also Italian wines carried to Hungary in tank trucks.

In our experiments the target molecule was the bovine mitochondrial DNA. We made the extraction of DNA using simple DNA extracting kit (Sigma Aldrich, Gen Elute). DNA yields and DNA concentrations was defined by usage of fluorometer (Qubit 2.0). As a further expectation, we had amplification of a short section, for the more sensitive detection, so we constructed primers for the CyB region of mitochondrial DNA. To detect the results of PCR more sensitively, we also tried the SYBR Green reactions.

Our researches revealed that the DNA, origins from milk, is detectable from wine, although the dilution of milk was 1:10000. According to our plan by continuing the experiments, the DNA sequence which is specific in cattle’s mitochondrial DNA will be identifiable in different levels of dilution both in the laboratory system modeled by us and in falsificated wines.



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