Polymerase chain reaction assay for verifying the labeling of meat and commercial meat products from game birds targeting specific sequences from the mitochondrial D-loop region

ArticleinPoultry Science 89(5):1021-1032 · May 2010with14 Reads
DOI: 10.3382/ps.2009-00217
Abstract
A PCR assay was developed for the identification of meats and commercial meat products from quail (Coturnix coturnix), pheasant (Phasianus colchicus), partridge (Alectoris spp.), guinea fowl (Numida meleagris), pigeon (Columba spp.), Eurasian woodcock (Scolopax rusticola), and song thrush (Turdus philomelos) based on oligonucleotide primers targeting specific sequences from the mitochondrial D-loop region. The primers designed generated specific fragments of 96, 100, 104, 106, 147, 127, and 154 bp in length for quail, pheasant, partridge, guinea fowl, pigeon, Eurasian woodcock, and song thrush tissues, respectively. The specificity of each primer pair was tested against DNA from various game and domestic species. In this work, satisfactory amplification was accomplished in the analysis of experimentally pasteurized (72°C for 30 min) and sterilized (121°C for 20 min) meats, as well as in commercial meat products from the target species. The technique was also applied to raw and sterilized muscular binary mixtures, with a detection limit of 0.1% (wt/wt) for each of the targeted species. The proposed PCR assay represents a rapid and straightforward method for the detection of possible mislabeling in game bird meat products.
    • "They used mt- DNA fragment to amplify specific DNA fragment for pork, lamb/mutton, chicken, ostrich meat, horse meat and beef, respectively. Furthermore, Rojas et al. (2010) reported that polymerase chain reaction assay can be used for verifying the labeling of meat and commercial meat products from game birds. This study amplified specific sequences from the mitochondrial D-loop region. "
    Full-text · Dataset · Mar 2016 · Food Control
    • "They used mt- DNA fragment to amplify specific DNA fragment for pork, lamb/mutton, chicken, ostrich meat, horse meat and beef, respectively. Furthermore, Rojas et al. (2010) reported that polymerase chain reaction assay can be used for verifying the labeling of meat and commercial meat products from game birds. This study amplified specific sequences from the mitochondrial D-loop region. "
    [Show abstract] [Hide abstract] ABSTRACT: By mixing with pork, beef adulteration is frequently found in the traditional market that very disturbing Moeslem community in Indonesia. This study was conducted to detect pork contamination in fresh and cooked beef using genetic marker mitochondrial DNA cytochrome b (mt-DNA Cyt b) by duplex-PCR. A total of twelve samples was used in this study consisting six fresh meat samples and six cooked meat samples, respectively. Those beef and pork were bought from animal slaughterhouse and a supermarket in Surakarta. Cooked samples were prepared by boiling the meats in hot water at 100oC for 30 minutes. We designed pork contamination in beef in the level of 0, 1, 5, 10, 25%, respectively. The DNA genome was extracted and polymerase chain reaction (PCR) was performed using species specific primer to isolate mt-DNA Cyt b gene from the samples. The results showed that the DNA genome was successfully extracted from pork, beef, and contaminated meat samples. In addition, visualization of duplex-PCR on 1.5% agarose gel was able to detect pork contamination in both fresh and cooked beef up to very small proportion (1%). The existence of pork in beef was indicated with the presence of specific 398 bp DNA band. It can be concluded, duplex-PCR of mt-DNA Cyt b gene was very sensitive in detection of pork contamination in fresh and cooked beef.
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    • "on of turkey, chicken, beef, pork and sheep meat in food products. TaqMan real-time PCR systems based on nucleotide sequence variation in D-loop and 12S rRNA mitochondrial genes were developed for the detection and quantification of DNA from chicken, turkey, duck, and goose material in highly processed industrial feed samples (Pegels et al., 2012). Rojas et al. (2010) , targeted the mitochondrial 12S rRNA gene for the identification of meats and commercial meat products from game birds, including quail, pheasant, partridge, guinea fowl, pigeon, Eurasian woodcock and song thrush. To our knowledge, there have been no reports on the use of realtime PCR method in seagull meat identification in meat produ"
    [Show abstract] [Hide abstract] ABSTRACT: The possibility of the adulteration of meat products with seagull meat disturbs people living in coastal cities. In order to eliminate the suspicions of consumers a sensitive and reliable method is needed for the detection of seagull meat. In order to identify and quantify seagull meat in meat mixtures a real-time polymerase chain reaction (PCR) assay, using species-specific primers and a TaqMan probe was designed on the mitochondrial NADH dehydrogenase subunit 2 gene.In addition, it was possible to detect the template DNA of seagull at the level of 100 pg without any cross-reactivity with non-target species (bovine, ovine, donkey, pork, horse, chicken, turkey, goose, duck). Also, the method was capable of detecting seagull meat at the level of 0.1% in raw and heat-treated test mixtures, prepared by mixing seagull meat with beef and chicken at different levels (0.01–10%). In conclusion, it can be suggested that the real-time PCR assay used in this research could be a rapid and sensitive method for the routine identification of seagull meat in raw or cooked meat products.
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