Abstract
Since game meat is more valuable and expensive than meat from domesticated animal species it is a potential target for adulteration. Analytical methods must allow the identification and quantification of meat species to be applicable for the detection of fraudulent labelling. We developed a real-time PCR assay for the authentication of sika deer (Cervus nippon) and products thereof. The primer/probe system amplifies a 71 bp fragment of the kappa-casein precursor gene. Since the target sequence contained only one sika deer-specific base, we introduced a deliberate base mismatch in the forward primer. The real-time PCR assay did not show cross-reactivity with 19 animal and 49 plant species tested. Low cross-reactivity was observed with red deer, fallow deer, reindeer and moose. However, with a ΔCt value of ≥11.79 between sika deer and the cross-reacting species, cross-reactivity will not affect the accuracy of the method. LOD and LOQ, determined by analysing serial dilutions of a DNA extract containing 1% (w/w) sika deer DNA in pig DNA, were 0.3% and 0.5%, respectively. The accuracy was evaluated by analysing DNA mixtures and DNA isolates from meat extract mixtures and meat mixtures. In general, recoveries were in the range from 70 to 130%.
Highlights
Commercial food products must not pose any health risks to consumers to comply with national and international food regulations[1,2]
Together with the respective reverse primer and probe, the forward primers were subjected to cross-reactivity tests with DNA isolates from red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus), alpine ibex (Capra ibex)
We aimed to develop a real-time polymerase chain reaction (PCR) assay for sika deer that can be used for authenticity testing of meat and meat products
Summary
Commercial food products must not pose any health risks to consumers to comply with national and international food regulations[1,2]. Sika deer and red deer differ in morphological characteristics, such as body size, shape of the antlers and pelage pattern Analytical methods must allow both the identification and quantification of meat species to be applicable to detect deer meat adulteration. We have already developed several real-time polymerase chain reaction (PCR) assays for the identification and quantification of deer meat in food products. One real-time PCR assay allows the identification and quantification of roe deer (Capreolus capreolus)[13], another one the determination of the sum of red deer (Cervus elaphus), fallow deer (Dama dama) and sika deer (Cervus nippon)[14]. We present a real-time PCR assay for the identification and quantification of sika deer in food products. Due to the constant copy number, single copy genes are better suitable for obtaining accurate quantitative results than mitochondrial DNA sequences[20]
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