Interlaboratory validation of a real-time PCR detection method for bovine- and ovine-derived material

Cai, Yicun; Wang, Qiang; He, Yihua; Pan, Liangwen

doi:10.1016/j.meatsci.2017.07.022

Cited by 5 publications

(4 citation statements)

References 21 publications

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“…In addition, we determined LOD 95% by applying a statistical model for calculating the probability of detection (POD) across laboratories. Since its introduction by Uhlig et al [27], this model has already been used several times to determine the sensitivity of real-time PCR assays [32][33][34][35][36][37]. Qualitative results obtained for the seven dilution steps of the DNA isolate from roe deer meat were used to determine the laboratory standard deviation σ L, and the LOD 95% for the median laboratory, as described previously [27].…”

Section: Lod 95% Derived From the Mixed Model For The Pod Curvementioning

confidence: 99%

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

Druml

Uhlig²,

Simon³

et al. 2021

Foods

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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 (LOD95%), 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.

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Section: Lod 95% Derived From the Mixed Model For The Pod Curvementioning

confidence: 99%

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

Druml

Uhlig²,

Simon³

et al. 2021

Foods

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“…Therefore, many countries and regions, including the European Union, requires that food and feedstuffs must be labelled with accurate and detailed information regarding the composition of all animal material [ 9 ]. Biological surveillance of the adulteration of animal materials is a major challenge for governmental agencies [ 10 ]. However, there are no widely acknowledged and recognized ISO (International Organization for Standardization) standards for the detection of animal material in foodstuffs.…”

Section: Introductionmentioning

confidence: 99%

“…In order to identify the species origin of animal materials in food and feedstuffs, DNA based PCR techniques have become important tools [ 10 , 11 ]. DNA contains species-specific information and indisputably describes biological diversity, and is an ideal target for molecular detection and identification of species-specific biological products [ 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

“…Many methods have been developed and applied to identify biological materials from specific animal species based on DNA analysis [ 11 , 14 – 20 ]. Presently, real-time PCR employing TaqMan probes is a well-established technique, and has been widely used due to its ease of operation, one-step procedure, good specificity, high accuracy, high efficiency, high sensitivity, and a low false-positive rate [ 10 , 13 , 21 – 26 ].…”

Section: Introductionmentioning

confidence: 99%

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Collaborative ring trial of two real-time PCR assays for the detection of porcine- and chicken-derived material in meat products

Wang

Cai

et al. 2018

PLoS ONE

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In this study, we describe an inter-laboratory collaborative ring trial validation of species-specific TaqMan real-time PCR assays for the detection of porcine- and chicken-derived materials in meat products. We comprehensively evaluated the performance of these assays in different environments and situations. This validation included the participation of thirteen laboratories across Europe and Asia. The results from the thirteen participating laboratories were analyzed to determine the specificity, accuracy, false positive rate, limit of detection (LOD), and probability of detection (POD) of the developed methods. These results indicated that the methods developed to detect porcine- and chicken-derived materials in meat products are robust and repeatable. The false positive and false negative rates were both 0%, and the LOD was determined to be five copies/reaction. The laboratory standard deviation (σL) was 0.30 for both detection methods, indicating that the developed methods are suitable for detection and identification of the porcine- and chicken-derived materials in meat products.

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

Wang

Cai

et al. 2017

Eur Food Res Technol

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Interlaboratory validation of a real-time PCR detection method for bovine- and ovine-derived material

Cited by 5 publications

References 21 publications

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

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

Collaborative ring trial of two real-time PCR assays for the detection of porcine- and chicken-derived material in meat products

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

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