European Food Research and Technology

, Volume 244, Issue 4, pp 767–774 | Cite as

Droplet digital PCR (ddPCR) method for the detection and quantification of goat and sheep derivatives in commercial meat products

  • Qiang Wang
  • Yicun Cai
  • Yuping He
  • Litao Yang
  • Jian Li
  • Liangwen Pan
Original Paper
  • 116 Downloads

Abstract

A highly precise, quantitative method based on the droplet digital polymerase chain reaction (ddPCR) technique was developed to identify and quantify the goat and sheep content in meat products. A formula for calculating raw meat weight based on DNA copy number was established. Exclusive specificity was verified using samples from 24 different animal species, and inclusive specificity between goat and sheep was tested using five different breeds for each species. The limit of detection and the limit of quantitation for both goat and sheep were 1 and 5 copies/μL, respectively, using a cloned plasmid containing goat- and sheep-specific target DNA fragments as calibrators. The accuracy and applicability of the method were verified using mixed powder samples with known proportions of goat and sheep meat, simulate meatball samples, and commercially available products, respectively. The results confirmed that the developed ddPCR methods are highly precise for identifying and quantifying the goat and sheep meat, indicating their potential applicability in future routine analyses.

Keywords

Droplet digital PCR Quantification Nuclear DNA Plasmid Meat product 

Notes

Acknowledgements

The authors are grateful to the reviewers for their careful corrections of the manuscript.

Compliance with ethical standards

Funding

Financial support from the Shanghai Science and Technology Commission Standard Special Fund (16DZ0501501), Shanghai Entry-Exit Food and Feed Safety Special Technology Service Platform Fund (17DZ2293700), Project funded by China Postdoctoral Science Foundation (2017M611628), Huaian Technical Fund (HAS201618) and Shanghai Entry-Exit Inspection and Quarantine Bureau of Science and Technology Plan Projects Fund (HK008-2017) are acknowledged with thanks.

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

Compliance with ethics requirements

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Qiang Wang
    • 1
  • Yicun Cai
    • 1
  • Yuping He
    • 1
  • Litao Yang
    • 2
  • Jian Li
    • 1
  • Liangwen Pan
    • 1
  1. 1.Technical Center for Animal, Plant and Food Inspection and QuarantineShanghai Entry-Exit Inspection and Quarantine Bureau of ChinaShanghaiChina
  2. 2.School of Life Science and BiotechnologyShanghai Jiao Tong UniversityShanghaiChina

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