For a rapid, specific and sensitive identification of cows', ewes' and goats' milk in mono-species Sicilian dairyproducts, species-specific duplex-PCR protocol was applied. DNA samples from blood and experimental cheesesof Sicilian autochthonous breeds were extracted to amplify the 12S rRNA (and part of 16S rRNA in case of Ovisaries) mitochondrial species-specific gene fragment. The use of species-specific primers for Bos taurus, Caprahircus and Ovis aries species, after electrophoresis on agarose gel, yielded fragments of 256 bp, 326 bp and172 bp, respectively. Amplification by duplex-PCR of DNA pools from two species showed detection thresholdsof 0.1% of “contaminant” DNA in each mixture. Finally, duplex-PCR assay was applied to experimental cheesesin order to detect the minimum threshold of DNA belonging to one species in cheese made with milk of twospecies. The results showed a sensitive threshold of 0.1% of ewes' milk in cows' and goats' cheeses, 0.1% of cows'milk in ewes' and goats' cheeses, and finally 0.1% of goats' milk in cows' and ewes' cheeses. The proposed assayrepresents a rapid and straightforward method of species traceability for the detections of adulteration inSicilian mono-species dairy products.
|Number of pages||6|
|Publication status||Published - 2016|
All Science Journal Classification (ASJC) codes
- Animal Science and Zoology