%0 Journal Article
%T Detection of Cow Milk in Water Buffalo Cheese by SYBR Green Real-Time PCR: Sensitivity Test on Governing Liquid Samples
%A Maria Feligini
%A Naima Alim
%A Ivan Bonizzi
%A Giuseppe Enne
%J Pakistan Journal of Nutrition
%D 2007
%I 
%X A real-time polymerase chain reaction (PCR) assay was developed to detect a bovine-specific mitochondrial DNA sequence in `buffalo` Mozzarella cheese by using primers targeting the cytochrome oxidase subunit 1 (COI) gene. Hot-start PCR, primer design, annealing and signal acquisition temperatures were exploited to obtain reliable analytical conditions which yielded a 134-bp amplicon from cow`s DNA only. Water buffalo`s DNA didn`t originate any amplification product. DNA isolated from blood was used to test primers` specificity and to construct a calibration curve in order to quantify bovine DNA concentration in governing liquid. The method was capable to detect low amounts of cow`s DNA in governing liquid samples. Due to low detection limit and fast, simple execution, the analytical protocol described in this work is suitable to become a common tool to detect the fraudulent addition of cow milk in water buffalo Mozzarella cheese.
%K Real-time PCR
%K sensitivity test
%K species identification
%K mitochondrial DNA
%K Mozzarella cheese
%U http://docsdrive.com/pdfs/ansinet/pjn/2007/94-98.pdf