Detection of Cow Milk in Water Buffalo Cheese by SYBR Green Real-Time PCR: Sensitivity Test on Governing Liquid Samples
Abstract
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.
Keywords:
Mitochondrial DNA, Mozzarella cheese, real-time PCR, sensitivity test, species identificationArticle Metrics
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Copyright (c) 2007 Asian Network for Scientific Information
This work is licensed under a Creative Commons Attribution 4.0 International License.
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.
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