title: Real-time PCR for quantitative detection of chamois (Rupicapra rupicapra) and pyrenean ibex (Capra pyrenaica) in meat mixtures.
abstract: A real-time quantitative polymerase chain reaction (PCR) technique was developed for the quantification of chamois and pyrenean ibex DNAs in meat mixtures by using a SYBR green detection platform. Two species-specific systems and a eukaryotic endogenous system were combined in the real-time PCR approach to quantify the target species. In the specific systems, a 133 base pair (bp) fragment of the 12S rRNA gene was amplified from chamois DNA, and an 88 bp fragment from the D-loop region was amplified from pyrenean ibex DNA. In the endogenous system, universal primers amplified a 141 bp fragment on the nuclear 18S rRNA gene from eukaryotic DNA. The threshold cycle values obtained with the 18S rRNA primers were used to normalize those obtained from chamois- or pyrenean ibex-specific systems, serving as endogenous control for the total content of PCR-amplifiable DNA in the sample. Analysis of experimental raw and heat-treated binary mixtures of chamois and pyrenean ibex meat in a swine meat matrix demonstrated the suitability of the assay for the detection and quantification of the target DNAs in the range of 0.1-0.8%, depending on the species and treatment of the meat samples.
authors: Universidad Complutense, Facultad de Veterinaria, Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, 28040 Madrid, Spain.
authors: Violeta
authors: V
authors: Fajardo
authors: Isabel
authors: I
authors: González
authors: Irene
authors: I
authors: Martín
authors: María
authors: M
authors: Rojas
authors: Pablo E
authors: PE
authors: Hernández
authors: Teresa
authors: T
authors: García
authors: Rosario
authors: R
authors: Martín
mesh_terms: D000818:Animals; D006041:Goats / Q000235:genetics*; D008460:Meat; D016133:Polymerase Chain Reaction / Q000379:methods*; D048608:Rupicapra / Q000235:genetics*; D012680:Sensitivity and Specificity