Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products

Sci Rep. 2018 May 8;8(1):7236. doi: 10.1038/s41598-018-25299-7.

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%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Caseins / analysis
  • Caseins / genetics*
  • DNA / analysis
  • DNA / genetics*
  • DNA / metabolism
  • DNA Primers / chemical synthesis
  • DNA Primers / metabolism
  • Deer
  • Food Labeling / ethics
  • Limit of Detection
  • Meat Products / analysis*
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / standards*
  • Sequence Alignment
  • Sequence Homology, Nucleic Acid
  • Species Specificity
  • Swine

Substances

  • Caseins
  • DNA Primers
  • DNA