Real-time PCR for quantitative detection of bovine tissues in food and feed.
A real-time PCR approach with the SYBR Green detection system has been developed for the quantitative detection of bovine tissues in food and feedstuffs. The method combines the use of bovine-specific primers, which amplify an 84-bp fragment of the mitochondrial 12S rRNA gene, and universal primers, which amplify a 140-bp fragment of the nuclear 18S rRNA gene from eukaryotic DNA. The 18S rRNA primers are used as endogenous controls for the total content of PCR-amplifiable DNA in the sample. The specificity of the primers was tested against 18 animal species, including mammals, birds, and fish, as well as 6 plant species. Analysis of experimental bovine tissues-oats mixtures demonstrated the suitability of the assay for the detection of bovine DNA in mixtures containing as low as 0.1% of bovine tissues. The performance of the method is not affected by severe heat treatment (up to 133 degrees C for 20 min at 300 kPa). The reported PCR assay could be very useful for detecting bovine-derived ingredients in raw and heat-treated food and feedstuffs.
Martín
Irene
I
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain.
García
Teresa
T
Fajardo
Violeta
V
Rojas
María
M
Hernández
Pablo E
PE
González
Isabel
I
Martín
Rosario
R
D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D002417:Cattle / Q000235:genetics*; D004247:DNA; D017931:DNA Primers / Q000032:analysis; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D006358:Hot Temperature; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D012335:RNA, Ribosomal; C075526:RNA, ribosomal, 12S; D004247:DNA