Accurate determination of meat mass fractions using DNA measurements for quantifying meat adulteration by digital PCR

Temisak, Sasithon; Thangsunan, Pattanapong; Boonnil, Jiranun; Yenchum, Watiporn; Hongthong, Kanjana; Boll, Heloísa Oss; Yata, Teerapong; Rios‐Solis, Leonardo; Morris, Phattaraporn

doi:10.1111/ijfs.15375

Cited by 9 publications

(2 citation statements)

References 43 publications

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“…Meat adulteration is a global issue that negatively affects trade on a national and international scale. The authentication of meat has attracted a lot of interest recently due to concerns about health, legal, religion, and economics (Chugunova 2020;El-Jalil et al 2020;Temisak et al 2021). For meat products to be of high food quality and safety, species discrimination procedures are essential.…”

Section: Discussionmentioning

confidence: 99%

Multiplex PCR Assay for Simultaneous Detection of Beef Meat Fraud with Different Species

2023

Int J Vet Sci

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Identification of species in animal-derived foods is a crucial component of its management. Legal, financial, religious, and public health considerations all apply to food adulteration, particularly in the case of beef meat. Food adulteration mostly includes replacement of low price ingredient in meat products to get unlawful higher benefits. To protect consumers from meat adulteration, various methods have been investigated. The DNA-based techniques are rapid, greater resistance to degradation, adequate for the detection of small amounts of DNA in complex handled food varieties. This work was designed to investigate different concentrations of beef meat adulterations with other meat types (equine, sheep, dog, and pork) using multiplex PCR. Fresh meat samples of different animal species cattle, donkey, pork and dog were collected for detection of their adulteration. Meat species samples were minced to make meat mixture for mimicking adulteration. The multiplex PCR assay for five meat species was run effectively, clarifying five unique PCR fragments. These PCR sections compared the particular sizes expected for the five designated species. The results showed successful amplification of the target cyt b gene sequences with the expected amplicon sizes (271pb) for cattle, (274pb) for sheep, and (808pb) for dog meat. Amplification of the target mt DNA, and 12S rRNA-tRNA Val gene sequences with the expected amplicon sizes (359pb) for equine, and (290bp) for pork's meat. The developed multiplex PCR assay was sensitive enough to detect 0.5% (w/w) adulterated meat under mixed matrices. It was concluded that the multiplex PCR could greatly minimize the cost for detection of meat adulteration.

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Section: Discussionmentioning

confidence: 99%

Multiplex PCR Assay for Simultaneous Detection of Beef Meat Fraud with Different Species

2023

Int J Vet Sci

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“…The majority of MBM testing uses the DNA amplification method Polymerase Chain Reaction (PCR), as it can detect even traces of fragmented DNA that survived the high temperatures and pressure from the rendering process. This method is equally suitable for the analysis of most meat-based food products for determining meat identity and food fraud [6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Development of Seven New dPCR Animal Species Assays and a Reference Material to Support Quantitative Ratio Measurements of Food and Feed Products

Griffiths,

McLaughlin,

Hall

et al. 2023

Foods

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Laboratory testing methods to confirm the identity of meat products and eliminate food fraud regularly rely on PCR amplification of extracted DNA, with most published assays detecting mitochondrial sequences, providing sensitive presence/absence results. By targeting single-copy nuclear targets instead, relative quantification measurements are achievable, providing additional information on the proportions of meat species detected. In this Methods paper, new assays for horse, donkey, duck, kangaroo, camel, water buffalo and crocodile have been developed to expand the range of species that can be quantified, and a previously published reference assay targeting the myostatin gene has been modified to include marsupials and reptiles. The accuracy of this ratio measurement approach was demonstrated using dPCR with mixtures of meat DNA down to 0.1%. However, the limit of detection (LOD) of this approach is not just determined by the assay targets, but by the samples themselves, with food or feed ingredients and processing impacting the DNA yield and integrity. In routine testing settings, the myostatin assay can provide multiple quality control roles, including monitoring the yield and purity of extracted DNA, identifying the presence of additional meats not detected by the suite of species-specific assays and potentially estimating a sample-specific LOD based on measured copy numbers of the myostatin target. In addition to the myostatin positive control assay, a synthetic DNA reference material (RM) has been designed, containing PCR targets for beef, pork, sheep, chicken, goat, kangaroo, horse, water buffalo and myostatin, to be used as a positive template control. The availability of standardised measurement methods and associated RMs significantly improves the reliability, comparability and transparency of laboratory testing, leading to greater confidence in results.

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MALDI-TOF MS Protein Profiling Combined with Multivariate Analysis for Identification and Quantitation of Beef Adulteration

Qiu

Huang

et al. 2022

Food Anal. Methods

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Accurate determination of meat mass fractions using DNA measurements for quantifying meat adulteration by digital PCR

Cited by 9 publications

References 43 publications

Multiplex PCR Assay for Simultaneous Detection of Beef Meat Fraud with Different Species

Multiplex PCR Assay for Simultaneous Detection of Beef Meat Fraud with Different Species

Development of Seven New dPCR Animal Species Assays and a Reference Material to Support Quantitative Ratio Measurements of Food and Feed Products

MALDI-TOF MS Protein Profiling Combined with Multivariate Analysis for Identification and Quantitation of Beef Adulteration

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