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

Overview

Journal Foods

Specialty Biotechnology

Date 2021 Nov 27

PMID 34828926

Citations 2

Authors

Barbara Druml

Steffen Uhlig

Kirsten Simon

Kirstin Frost

Karina Hettwer

Margit Cichna-Markl

Rupert Hochegger

Affiliations

Soon will be listed here.

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 % (/) 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% (/) 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), 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.

Citing Articles

A Novel Normalized Quantitative Real-Time PCR Approach for Ensuring Roe Deer () Meat Authenticity in Game Meat Foods.

Adenuga B, Biltes R, Villa C, Costa J, Spychaj A, Montowska M Foods. 2024; 13(23).

PMID: 39682800 PMC: 11640146. DOI: 10.3390/foods13233728.

Techniques for Food Authentication: Trends and Emerging Approaches.

Cichna-Markl M, Mafra I Foods. 2023; 12(6).

PMID: 36981061 PMC: 10048066. DOI: 10.3390/foods12061134.

References

Cai Y, Wang Q, He Y, Pan L . Interlaboratory validation of a real-time PCR detection method for bovine- and ovine-derived material. Meat Sci. 2017; 134:119-123. DOI: 10.1016/j.meatsci.2017.07.022. View

Kaltenbrunner M, Mayer W, Kerkhoff K, Epp R, Ruggeberg H, Hochegger R . Differentiation between wild boar and domestic pig in food by targeting two gene loci by real-time PCR. Sci Rep. 2019; 9(1):9221. PMC: 6593100. DOI: 10.1038/s41598-019-45564-7. View

Druml B, Mayer W, Cichna-Markl M, Hochegger R . Development and validation of a TaqMan real-time PCR assay for the identification and quantification of roe deer (Capreolus capreolus) in food to detect food adulteration. Food Chem. 2015; 178:319-26. DOI: 10.1016/j.foodchem.2015.01.003. View

Fajardo V, Gonzalez I, Martin I, Rojas M, Hernandez P, Garci A T . Differentiation of European wild boar (Sus scrofa scrofa) and domestic swine (Sus scrofa domestica) meats by PCR analysis targeting the mitochondrial D-loop and the nuclear melanocortin receptor 1 (MC1R) genes. Meat Sci. 2011; 78(3):314-22. DOI: 10.1016/j.meatsci.2007.06.018. View

Druml B, Grandits S, Mayer W, Hochegger R, Cichna-Markl M . Authenticity control of game meat products--a single method to detect and quantify adulteration of fallow deer (Dama dama), red deer (Cervus elaphus) and sika deer (Cervus nippon) by real-time PCR. Food Chem. 2014; 170:508-17. DOI: 10.1016/j.foodchem.2014.08.048. View

Ballin N . Authentication of meat and meat products. Meat Sci. 2010; 86(3):577-87. DOI: 10.1016/j.meatsci.2010.06.001. View

Kaltenbrunner M, Mayer W, Kerkhoff K, Epp R, Ruggeberg H, Hochegger R . Applicability of a duplex and four singleplex real-time PCR assays for the qualitative and quantitative determination of wild boar and domestic pig meat in processed food products. Sci Rep. 2020; 10(1):17243. PMC: 7560752. DOI: 10.1038/s41598-020-72655-7. View

Kaltenbrunner M, Hochegger R, Cichna-Markl M . Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products. Sci Rep. 2018; 8(1):7236. PMC: 5940659. DOI: 10.1038/s41598-018-25299-7. View

Gondim C, Junqueira R, de Souza S, Callao M, Ruisanchez I . Determining performance parameters in qualitative multivariate methods using probability of detection (POD) curves. Case study: Two common milk adulterants. Talanta. 2017; 168:23-30. DOI: 10.1016/j.talanta.2016.12.065. View

10.

Hird H, Chisholm J, Sanchez A, Hernandez M, Goodier R, Schneede K . Effect of heat and pressure processing on DNA fragmentation and implications for the detection of meat using a real-time polymerase chain reaction. Food Addit Contam. 2006; 23(7):645-50. DOI: 10.1080/02652030600603041. View

11.

Ballin N, Vogensen F, Karlsson A . Species determination - Can we detect and quantify meat adulteration?. Meat Sci. 2010; 83(2):165-74. DOI: 10.1016/j.meatsci.2009.06.003. View

12.

Fumiere O, Dubois M, Baeten V, von Holst C, Berben G . Effective PCR detection of animal species in highly processed animal byproducts and compound feeds. Anal Bioanal Chem. 2006; 385(6):1045-54. DOI: 10.1007/s00216-006-0533-z. View

13.

DAmato M, Alechine E, Cloete K, Davison S, Corach D . Where is the game? Wild meat products authentication in South Africa: a case study. Investig Genet. 2013; 4(1):6. PMC: 3621286. DOI: 10.1186/2041-2223-4-6. View

14.

Fraiture M, Papazova N, Roosens N . DNA walking strategy to identify unauthorized genetically modified bacteria in microbial fermentation products. Int J Food Microbiol. 2020; 337:108913. DOI: 10.1016/j.ijfoodmicro.2020.108913. View

15.

Rojas M, Gonzalez I, Pavon M, Pegels N, Lago A, Hernandez P . Novel TaqMan real-time polymerase chain reaction assay for verifying the authenticity of meat and commercial meat products from game birds. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2010; 27(6):749-63. DOI: 10.1080/19440040903503070. View

16.

Kaltenbrunner M, Hochegger R, Cichna-Markl M . Development and validation of a fallow deer (Dama dama)-specific TaqMan real-time PCR assay for the detection of food adulteration. Food Chem. 2017; 243:82-90. DOI: 10.1016/j.foodchem.2017.09.087. View

17.

Hoffman L, Wiklund E . Game and venison - meat for the modern consumer. Meat Sci. 2011; 74(1):197-208. DOI: 10.1016/j.meatsci.2006.04.005. View

18.

Kaltenbrunner M, Hochegger R, Cichna-Markl M . Tetraplex real-time PCR assay for the simultaneous identification and quantification of roe deer, red deer, fallow deer and sika deer for deer meat authentication. Food Chem. 2018; 269:486-494. DOI: 10.1016/j.foodchem.2018.07.023. View

19.

Fajardo V, Gonzalez I, Lopez-Calleja I, Martin I, Rojas M, Hernandez P . Identification of meats from red deer (Cervus elaphus), fallow deer (Dama dama), and roe deer (Capreolus capreolus) using polymerase chain reaction targeting specific sequences from the mitochondrial 12S rRNA gene. Meat Sci. 2011; 76(2):234-40. DOI: 10.1016/j.meatsci.2006.11.004. View

20.

Wang Q, Cai Y, He Y, Yang L, Pan L . Collaborative ring trial of two real-time PCR assays for the detection of porcine- and chicken-derived material in meat products. PLoS One. 2018; 13(10):e0206609. PMC: 6205609. DOI: 10.1371/journal.pone.0206609. View