DNA条形码COI序列在常见肉类鉴别中的应用研究

Cytochrome C Oxidase Subunit I sequence as a DNA Barcode to Identify Common Meat Species

摘要:

为了对常见的4种肉类及相关肉制品进行掺假鉴定，判别与产品标签是否相符，本研究以COI基因为靶基因，建立了4种动物源性食品DNA条形码鉴别技术。分别提取牛、羊、猪、鸭四大物种的基因组DNA为模板，以其COI基因的保守序列区设计6对通用引物，结合文献报道及数据库提供的7对通用引物进行PCR扩增，并将测序结果提交GenBank数据库Blast比对，评价不同DNA条形码的检测鉴别能力。筛选出COI-A为最优序列，在4个物种中扩增效率100%。对抽检的20个批次的肉加工品样品进行检测，鉴定结果约有90%的样品与产品标签标示的成分相符。其中1个批次的牛丸制品因肉类成分含量低未扩增成功，1个批次的牛丸制品检出鸭源成分，判定掺假。DNA条形码技术快速有效，本研究筛选的COI-A序列可直接用于牛、羊、猪、鸭及其肉制品的鉴定，并为其它常见动物源性食品的种类鉴定提供一定参考依据。

Abstract:

To detect adulteration in four common meat species and related meat products, and determine if the products are consistent with the label description, a DNA barcoding technique was established in this study. The cytochrome c oxidase subunit I (COI) gene was used as the target gene to identify four types of animal-derived foods. Genomic DNA from cattle, sheep, pig, and duck was extracted and used as template and six pairs of universal primers were designed based on the conserved regions of the COI gene. Polymerase chain reaction (PCR) amplification was performed using these six pairs of universal primers as well as seven pairs of universal primers obtained from the literature. The sequencing results were aligned using basic local alignment search tool (BLAST) analysis of the GenBank database, to evaluate the identification and detection capabilities of each DNA barcode. The COI-A sequence was found to be optimal, showing 100% amplification efficiency in four species. A total of 20 batches of meat product samples were analyzed using the primer COI-A and approximately 90% samples were found to be consistent with their label descriptions. Amplification failed for one batch of beef-ball sample due to low meat content, while duck-derived material was detected in a second batch, indicating adulteration. DNA barcoding technique is rapid and effective, and the COI-A sequence selected in this study can be applied to directly identify beef, lamb, pork, and duck as well as related meat products. The results of this study provide a reference for species identification of other common animal-derived foods.