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用于检测和定量狍以检测食品掺假的实时聚合酶链反应检测法——涉及奥地利、德国和瑞士实验室的实验室间验证

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.

作者信息

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

机构信息

Department of Molecular Biology and Microbiology, Institute for Food Safety Vienna, Austrian Agency for Health and Food Safety (AGES), Spargelfeldstraße 191, 1220 Vienna, Austria.

Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna, Währinger Straße 38, 1090 Vienna, Austria.

出版信息

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

DOI:10.3390/foods10112645
PMID:34828926
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8623729/
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.

摘要

野味肉类产品特别容易被用更便宜的肉类品种替代野味肉来掺假。最近,我们提出了一种实时聚合酶链反应(PCR)检测方法,用于食品中狍子的鉴定和定量。通过对DNA分离物除进行狍子实时PCR检测外,还进行参考实时PCR检测,相对地实现了狍子含量以%(/)为单位的定量。为了协调欧盟成员国食品认证的分析方法,狍子实时PCR检测方法在一项实验室间循环试验中进行了测试,该试验包括来自奥地利、德国和瑞士的14个实验室。参与实验室从一份猪肉中含有24.8%(/)狍子肉的混合肉、狍子肉以及12份未披露狍子含量的肉类样品中获得了DNA分离物的等分试样。性能特征包括扩增效率、检测限(LOD)、重复性、再现性以及定量结果的准确性。发现该实时PCR检测方法的相对再现性标准偏差在13.35%至25.08%之间(去除异常值后),回收率在84.4%至114.3%之间,适用于生肉样品中狍子的检测和定量,以检测食品掺假。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876e/8623729/c208f30f2dbb/foods-10-02645-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876e/8623729/b8ad445a440b/foods-10-02645-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876e/8623729/9eae74f27586/foods-10-02645-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876e/8623729/c208f30f2dbb/foods-10-02645-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876e/8623729/b8ad445a440b/foods-10-02645-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876e/8623729/9eae74f27586/foods-10-02645-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876e/8623729/c208f30f2dbb/foods-10-02645-g003.jpg

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