Abstract
Verifying the authenticity of food products is essential due to the recent increase in counterfeit meat-containing food products. The existing methods of detection have a number of disadvantages. Therefore, simple, cheap, and sensitive methods for detecting various types of meat are required. In this study, we propose a rapid full-cycle technique to control the chicken or pig adulteration of meat products, including 3 min of crude DNA extraction, 20 min of recombinase polymerase amplification (RPA) at 39 °C, and 10 min of lateral flow assay (LFA) detection. The cytochrome B gene was used in the developed RPA-based test for chicken and pig identification. The selected primers provided specific RPA without DNA nuclease and an additional oligonucleotide probe. As a result, RPA–LFA, based on designed fluorescein- and biotin-labeled primers, detected up to 0.2 pg total DNA per μL, which provided up to 0.001% w/w identification of the target meat component in the composite meat. The RPA–LFA of the chicken and pig meat identification was successfully applied to processed meat products and to meat after heating. The results were confirmed by real-time PCR. Ultimately, the developed analysis is specific and enables the detection of pork and chicken impurities with high accuracy in raw and processed meat mixtures. The proposed rapid full-cycle technique could be adopted for the authentication of other meat products.
Highlights
In modern society, falsifying the composition of food products by violating the declared recipe has become a serious problem [1,2,3,4]
We propose a rapid full-cycle technique to control the chicken or pig adulteration of meat products, including 3 min of crude DNA extraction, 20 min of recombinase polymerase amplification (RPA) at 39 ◦C, and 10 min of lateral flow assay (LFA) detection
All forward primers were modified by biotin at the 5 terminal, and reverse primers were modified by FAM at the 5 terminal
Summary
In modern society, falsifying the composition of food products by violating the declared recipe has become a serious problem [1,2,3,4]. The variety of methods used to solve these problems is extremely wide and includes the identification of specific biomarkers obtained by electrophoresis and chromatography [17,18,19] and assessment of the compositional characteristics of raw materials via microscopy and spectroscopy [20,21,22,23]. These methods are extremely laborious, characterized by low productivity, and require highly qualified personnel and expensive equipment that is available only to a limited number of centralized laboratories. PCR is suitable for confirming conclusions, but simpler and more mobile methods of initial screening remain to be determined
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