Detection of meat adulteration: Use of efficient and routine-suited multiplex polymerase chain reaction-based methods for species authentication and quantification in meat products
2018
Zdeňková, Kamila | Akhatova, Diliara | Fialová, Eliška | Krupa, Ondřej | Kubica, Lukáš | Lencová, Simona | Demnerová, Kateřina
The increase in the extent of meat adulteration is the reason for a need for an effective method for authentication of meat products. DNA-based polymerase chain reaction (PCR) is a well suited alternative for this purpose. Furthermore, the method facilitates quantification of animal DNA in meat products based on the correlation between target copy amounts and cycle numbers in quantitative PCR. We designed and experimentally verified PCR primer systems for identification of beef, pork, horse and poultry (chicken, turkey) meat. Mitochondrial and chromosomal markers were used. The mitochondrial cytochrome b gene was used as a marker for qualitative multiplex endpoint PCR and single- copy chromosomal genes (cyclic-GMP-phosphodiesterase gene for cattle, beta-actin gene for pig, interleukin-2 gene for chicken, myostatin gene for mammals and poultry) were used for multiplex quantitative PCR analyses. The reliability of both methods was confirmed by analysing of mixed samples prepared with or without heat treatment. The methods were then applied to 14 commercially available products typical for the Czech Republic, including sausages or salami. Discrepancies were observed between the DNA analysis and the meat content declared for the tested products, as two of the samples did not correspond to qualitative requirements and other four failed to meet quantitative requirements. The proposed PCR-based methodology was shown to be useful for the disclosure of meat adulteration.
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