基于COI序列的DNA微条形码技术鉴别熟肉制品中11种肉掺假的研究

建立并优化了基于COI序列的DNA微条形码技术(mini-barcoding)检测熟肉制品中11种常见肉类掺假的方法.样品经超声与真空冷冻干燥处理,提取DNA模板和PCR扩增后,目标扩增物经切胶纯化后进行克隆测序,并将测序结果提交GenBank数据库Blast比对.筛选出适合猪、牛、羊、鸡、鸭、鸽子、马、驴、鹅、兔、鼠...

Identification of 11 Adulterated Meat Species in Cooked Meat Products Using DNA Mini-barcoding Methods Based on Cytochrome C Oxidase Subunit I Sequence(COI)

A DNA min-barcoding method with cytochrome C oxidase subunit I sequence(COI)was developed to test 11 adulterated meat species in cooked meat products.DNA extraction and amplification of the mini-barcode region were carried out after samples were treated with ultrasonication and vacuum freeze drying.PCR products were purified with DNA purification kit and analyzed by clone sequencing.The sequencing results were submitted in GenBank using Blast.The primer set COI-A was found to be optimal,showing 100%amplification efficiency in 11 cooked meat species(including pork,beef,mutton,chicken,duck,pigeon meat,horsemeat,ass meat,goose,rabbit meat and rat meat)with optimized PCR procedure.18 meat adulteration models were founded to test the minimum detected adulteration rate by cheap meat species including pork,chicken,duck,horsemeat,ass meat,and rat meat,and results showed that the minimum detected adulteration rates of beef and mutton added by the 6 cheap species were 5%with chicken was 10%,and the rates of pigeon,goose and rabbit meat were 10%.Using this method to test 30 cooked meat products,18 were found to be adulterated.The method was simple,sensitive and reproducible which would be suitable for the detection of 11 adulterated meat species in cooked meat products.