Real-Time PCR Assay for the Detection and Quantification of Roe Deer to Detect Food Adulteration-Interlaboratory Validation Involving Laboratories in Austria, Germany, and Switzerland

Barbara Druml; Steffen Uhlig; Kirsten Simon; Kirstin Frost; Karina Hettwer; Margit Cichna-Markl; Rupert Hochegger

doi:10.3390/foods10112645

Real-Time PCR Assay for the Detection and Quantification of Roe Deer to Detect Food Adulteration-Interlaboratory Validation Involving Laboratories in Austria, Germany, and Switzerland

Foods. 2021 Nov 1;10(11):2645. doi: 10.3390/foods10112645.

Authors

Barbara Druml^{1

2}, Steffen Uhlig³, Kirsten Simon³, Kirstin Frost³, Karina Hettwer³, Margit Cichna-Markl², Rupert Hochegger¹

Affiliations

¹ Department of Molecular Biology and Microbiology, Institute for Food Safety Vienna, Austrian Agency for Health and Food Safety (AGES), Spargelfeldstraße 191, 1220 Vienna, Austria.
² Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna, Währinger Straße 38, 1090 Vienna, Austria.
³ QuoData GmbH, Prellerstraße 14, 01309 Dresden, Germany.

Abstract

Game meat products are particularly prone to be adulterated by replacing game meat with cheaper meat species. Recently, we have presented a real-time polymerase chain reaction (PCR) assay for the identification and quantification of roe deer in food. Quantification of the roe deer content in % (w/w) was achieved relatively by subjecting the DNA isolates to a reference real-time PCR assay in addition to the real-time PCR assay for roe deer. Aiming at harmonizing analytical methods for food authentication across EU Member States, the real-time PCR assay for roe deer has been tested in an interlaboratory ring trial including 14 laboratories from Austria, Germany, and Switzerland. Participating laboratories obtained aliquots of DNA isolates from a meat mixture containing 24.8% (w/w) roe deer in pork, roe deer meat, and 12 meat samples whose roe deer content was not disclosed. Performance characteristics included amplification efficiency, level of detection (LOD_95%), repeatability, reproducibility, and accuracy of quantitative results. With a relative reproducibility standard deviation ranging from 13.35 to 25.08% (after outlier removal) and recoveries ranging from 84.4 to 114.3%, the real-time PCR assay was found to be applicable for the detection and quantification of roe deer in raw meat samples to detect food adulteration.

Keywords: detection; food authentication; game meat; interlaboratory ring trial; probability of detection; quantification; real-time PCR; roe deer; validation.