Real-time PCR based on single-copy housekeeping genes for quantitative detection of goat meat adulteration with pork

Adulteration of goat meat with cheaper meat such as pork has been frequently found. Conventional PCR methods to distinguish goat meat from adulteration are qualitative, which easily produce false-positive results due to contamination but not adulteration. To address this problem, real-time PCR based on single-copy housekeeping genes encoding replication protein A1 was developed for goat meat and pork. By calculating the Ct ratio of goat meat/pork, the goat meat content in a suspected adulteration could be deduced by a good linear correlation (R-2 = 0.9868) in a range from 5% to 80%. Analysis of the simulated samples of goat meat showed high accuracy with recoveries of 104.91% and 105.00% for the goat meat contents of 40% and 60%, respectively, and coefficient of variations were as low as 9.24% and 9.09%. Thus, the developed assay supplied a useful tool for food regulation authorities to achieve quantitative authentication of goat meat.