Real-time PCR for quantitative detection of chamois (Rupicapra rupicapra) and pyrenean ibex (Capra pyrenaica) in meat mixtures.
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.
Fajardo
Violeta
V
Universidad Complutense, Facultad de Veterinaria, Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, 28040 Madrid, Spain.
González
Isabel
I
Martín
Irene
I
Rojas
María
M
Hernández
Pablo E
PE
García
Teresa
T
Martín
Rosario
R
D000818:Animals; D006041:Goats / Q000235:genetics*; D008460:Meat; D016133:Polymerase Chain Reaction / Q000379:methods*; D048608:Rupicapra / Q000235:genetics*; D012680:Sensitivity and Specificity