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

Departamento de Nutrición, Bromatología y Tecnología de los Alimentos Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain.
Poultry Science (Impact Factor: 1.54). 05/2010; 89(5):1021-1032. 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 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.

1 Follower
  • Source
    [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.
    Food Control 11/2013; 34(1):47–49. DOI:10.1016/j.foodcont.2013.04.006 · 2.82 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Economical, religious, and health reasons demand an accurate control of food in order to protect consumers from falsely labeled products. Meat in particular is easily susceptible to fraudulent labeling, mainly through contamination with species of lower value. New methods and protocols for rapid, sensitive and reliable identification of extraneous species in food are therefore required. The miniaturization and optimization of analytical methodologies are powerful tools in this direction, especially when connected to Lab-on-a-chip (LOC) microdevices. LOCs possess many advantages, such as the reduction of the analysis cost, the possibility to save time and labor, the easiness of use and not last, the possibility to bring a complex technique out of the laboratory. Here we present a new concept for the food quality control, i.e. the use of LOC for the detection of exogenous DNA in meat via on-chip PCR in real-time. LOC surfaces were treated with different coatings in order to optimize the DNA extraction directly from meat homogenates (bovine, pork, horse). On the same LOC used for DNA purification, we set up the on-chip PCR with real-time detection. Over 1,000 beef genomes, up to 0.01 horse or pork genomes were successfully detected in binary mixtures of pre-purified DNA and similarly, up to 0.01 % parts of exogenous meat were detected in binary mixtures of meat homogenates. The successful on-chip detection of exogenous DNA is a promising step toward the production of an effective microdevice for rapid, sensitive, and reliable identification of meat adulteration.
    02/2013; DOI:10.1007/s12668-013-0080-y
  • [Show abstract] [Hide abstract]
    ABSTRACT: Food authentication and quality control requires sensitive species-specific identification and quantification of DNA from unknown sources. An accurate and reliable multiplex TaqMan® real-time quantitative PCR (qPCR) method modified from Köppel et al. (2009) based on the specific sites of nuclear DNA has been developed to identify duck, pig and chicken DNA in blood curds. Total DNA in blood curd samples was extracted using three methods: TIANamp® Blood DNA Kit, TIANamp® Genomic DNA Kit and a modified phenol/chloroform extraction method. The concentration and purity of DNA was compared regarding DNA yields and purity. A modified phenol/chloroform extraction method and the TIANamp® Genomic DNA Kit gave improved extraction efficiencies compared with the TIANamp® Blood DNA Kit. The limit of detection (LOD) of the assay was 0.15 ng for each target species (1: 103 dilution). Analysis of experimental mixtures demonstrated that the sensitivity of the assay was 1% for each species analyzed. This system proved its accuracy, precision and applicability for the examination of different types of animal blood in blood curd products of the type presently on the market.
    Food Research International 06/2014; 60. DOI:10.1016/j.foodres.2014.01.047 · 3.05 Impact Factor