Skip to main content
Log in

Digital duplex versus real-time PCR for the determination of meat proportions from sausages containing pork and beef

  • Original Paper
  • Published:
European Food Research and Technology Aims and scope Submit manuscript

Abstract

According to Swiss law, sausages claiming to be made of veal have to contain at least 50% veal. To control the meat proportion of such products, control laboratories use real-time PCR. The measurement uncertainty of this method is at 30%. As a consequence, only extreme fraud can be reliably detected. To analyse sausages for their beef content with lower measurement uncertainty, a duplex droplet digital PCR was developed. Interlaboratory conversion factors were determined to enable weight-to-weight determination using values gained by real-time PCR and droplet digital PCR. Precision and accuracy were investigated examining reference sausages and sausages from the market. Comparing with real-time PCR, results from digital PCR showed a superior interlaboratory measurement uncertainty of 10% and will enable food control laboratories to determine also minor fraud.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Swiss Food Legislation Verordnung über Lebensmittel tierischer Herkunft; 16. Dezember 2016; Art. 9 Abs. 5

  2. Sawyer J, Wood C, Shanahan D, Gout S, McDowell D (2003) Real-time PCR for quantitative meat species testing. Food Control 14:579–583

    Article  CAS  Google Scholar 

  3. Brodmann P, Moor D (2003) Sensitive and semi-quantitative TaqMan® real-time polymerase chain reaction systems for the detection of beef (Bos taurus) and the detection of the family Mammalia in food and feed. Meat Sci 65:599–607

    Article  CAS  PubMed  Google Scholar 

  4. Lopez-Andreo M, Garrido-Pertierra LL, Prieto A, Puyet I A (2005) Identification and quantitation of species in complex DNA mixtures by real-time polymerase chain reaction. Anal Biochem 339:73–82

    Article  CAS  PubMed  Google Scholar 

  5. Laube I, Zagon J, Broll H (2007) Quantitative determination of commercially relevant species in foods by real-time PCR. International Journal of Food Science Technology 42:336–341

    Article  CAS  Google Scholar 

  6. Eugster A, Ruf J, Rentsch J, Hübner P, Köppel R (2008) Quantification of beef and pork fraction in sausages by real-time PCR analysis: results of an interlaboratory trial. European Food Research Technology 227(1):17–20

    Article  CAS  Google Scholar 

  7. Köppel R, Ruf J, Zimmerli F, Breitenmoser A (2008) Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, chicken and turkey. European Food Research Technology 227(4):1199–1203

    Article  CAS  Google Scholar 

  8. Eugster A, Ruf J, Rentsch J, Köppel R (2009) Quantification of beef, pork, chicken and turkey proportions in sausages: use of matrix-adapted standards and comparison of single versus multiplex PCR in a interlaboratory trial. European Food Research Technology 230:55–61

    Article  CAS  Google Scholar 

  9. Jonker KM, Tilburg JHC, Hägele GH, De Boer E (2008) Species identification in meat products using real-time. PCR Food Additives Contaminants 25(5):527–533

    Article  CAS  PubMed  Google Scholar 

  10. Köppel R, Ruf J, Rentsch J (2011) Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, horse and sheep. Eur Food Res Technol 232:151–155

    Article  CAS  Google Scholar 

  11. Cai Y, Li X, Lv R, Yang J, Li J, He Y, Pan L (2014) Quantitative Analysis of Pork and Chicken Products by Droplet Digital PCR. BioMed Research International Volume 2014 (2014), Article ID 810209, 6 pages

  12. Floren C, Wiedemann I, Brenig B, Schütz E, Beck J (2015) Species identification and quantification in meat and meat products using Droplet. Digital PCR (ddPCR. Food Chem 173:1054–1058

    Article  CAS  PubMed  Google Scholar 

  13. Shehata R, Li RH,J, Chen S, Helen Redda H, Cheng S, Li TN,H, Hanner WK R (2017) Droplet digital polymerase chain reaction (ddPCR) assays integrated with an internal control for quantification of bovine, porcine, chicken and turkey species in food and feed. PLOS one. doi.https://doi.org/10.1371/journal.pone.0182872

  14. Wang Q, Cai Y, He Y, Yang L, Li J, Pan L (2018) Droplet digital PCR (ddPCR) method for the detection an quantification of goat and sheep derivatives in commercial meat products. Eur Food Res Technol 244:767–774

    Article  CAS  Google Scholar 

  15. Uhlig S, Baldauf H, Simon K, Methodenringversuch “All Meat” zum quantitativen Nachweis von DNA der Tierarten Huhn, Pute, Rind und Schwein mittels Tetraplex real-time PCR, Quodata, in preparation

  16. Gerdes L, Iwobi A, Busch U, Pecoraro S (2016) Optimization of digital droplet polymerase chain reaction for quanitifcation of genetically modified organism. Biomolecular detection Quantification 7:9–20

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank the cantonal laboratory of Zürich, the Official Food Control Authority of the Canton St.Gallen, Cantonal Office of Consumer Protection Bern, Switzerland and AGES CC Biochemie Vienna, Austria, for providing sample material and resources for this work.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to René Köppel.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Compliance with ethics requirements

This article does not contain any studies with human or living animal subjects.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Köppel, R., Ganeshan, A., van Velsen, F. et al. Digital duplex versus real-time PCR for the determination of meat proportions from sausages containing pork and beef. Eur Food Res Technol 245, 151–157 (2019). https://doi.org/10.1007/s00217-018-3147-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00217-018-3147-8

Keywords

Navigation