Real-time PCR Detection for Buffalo(Bubalus bubalus) Ingredients in Dairy Products

Dairy product authenticity is presently a subject of great concern to food authorities,as the incorrect labelling of foodstuffs can present a commercial fraud.Many consumers are concerned about dairy products they eat and accurate labelling is important to inform consumer choice.Thus,the detection of milk species is important in cheese making,not only with regards to accurate consumer information and legal aspects(concerned with labeling and guarantee requirements) but also to medical requirements,food allergies or religious practices.Specific primers and Taq Man-MGB probe were designed according to highly conserved sequence of mitochondrial cytochrome b(Cytb) gene of buffalo(Bubalus bubalus) to develop a Real-time fluorescence polymerase chain reaction(Real-time PCR) assay for rapid detection of buffalo ingrediens.The primers and probes could specific identify 5 kinds of buffalo with 47 kinds of samples.The detection limit of the method was 0.001 ng/μL DNA concentration and 0.01 %(W/W) buffalo milk.The commercial samples were detected by Real-time PCR and the detection results showed that buffalo ingredients were detected well.All the results confirm that the Real-time PCR assay is a sensitive and specific method for the detection of buffalo materials in dairy products.