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
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 degrees C for 30 min) and sterilized (121 degrees 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.
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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.Food Control 11/2013; 34(1):47–49. DOI:10.1016/j.foodcont.2013.04.006 · 2.82 Impact Factor
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ABSTRACT: Two PCR assays for the identification of partridge meat (red-legged partridge, chukar partridge, barbary partridge, and gray partridge species) and the specific identification of red-legged partridge meat products were developed based on species-specific primers targeting the 12S ribosomal RNA mitochondrial gene. Moreover, various PCR techniques based on the use of random amplified polymorphic DNA markers and nuclear growth hormone and rhodopsin genes were tested to find a method for the differentiation between pure and hybrid red-legged partridges. Among these techniques, the PCR method based on the amplification and sequencing of a nuclear rhodopsin gene fragment was selected as a suitable tool for the discrimination among meats from pure and hybrid red-legged partridge individuals. The PCR assays reported in this work could be useful in inspection programs to verify the correct labeling of raw and heat-treated partridge meat products.Poultry Science 01/2011; 90(1):211-222. DOI:10.3382/ps.2010-00895 · 1.67 Impact Factor
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ABSTRACT: A species-specific real-time polymerase chain reaction (PCR) assay using TaqMan (R) probes has been developed for the identification of meat and meat products from common pigeon (Columba livia), woodpigeon (Columba palumbus) and stock pigeon (Columba oenas). The method combines the use of species-specific primers and TaqMan (R) probes that amplify small fragments (amplicons < 200 base pairs) of the mitochondrial 12S rRNA gene, and an endogenous control primer pair that amplifies a 141 bp fragment of the nuclear 18S rRNA gene from eukaryotic DNA. Analysis of experimental raw and heat-treated binary mixtures as well as of commercial meat products from the target species, demonstrated the suitability of the assay for the detection of the target DNAs. The PCR assay reported in this work could be useful in inspection programs to verify the correct labelling of raw and heat-treated pigeon meat products.Food Control 02/2012; 23(2):369-376. DOI:10.1016/j.foodcont.2011.07.034 · 2.82 Impact Factor