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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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Meat products often consist of meat from multiple animal species, and inaccurate food product adulteration and mislabeling can negatively affect consumers. Therefore, a cost-effective and reliable method for identification and quantification of animal species in meat products is required. In this study, we developed a duplex droplet digital PCR (dddPCR) detection and quantification system to simultaneously identify and quantify the source of meat in samples containing a mixture of beef (Bos taurus) and pork (Sus scrofa) in a single digital PCR reaction tube. Mixed meat samples of known composition were used to test the accuracy and applicability of this method. The limit of detection (LOD) and the limit of quantification (LOQ) of this detection and quantification system were also identified. We conclude that our dddPCR detection and quantification system is suitable for quality control and routine analyses of meat products. [ABSTRACT FROM AUTHOR]

In this work we performed interlaboratory validation of a Taqman real-time PCR method for the identification of bovine and ovine material. The Bos taurus beta-actin gene (ACTB) and Ovis aries prolactin receptor gene (PRLR) were selected as the bovine and ovine species-specific amplifying target sequences, and primers and TaqMan probes were designed accordingly. The precision, efficiency, false positive rate, limit of detection (LOD 95% ) and probability of detection (POD) were determined, and the results demonstrated that both bovine and ovine detection methods performed well. The high homogeneity of the results indicates that the detection methods are suitable for a wide range of applications, and the tools developed herein could be applied by official and third-party detection institution to maintain quality in the food and feedstuff industries. [ABSTRACT FROM AUTHOR]