摘要
采用DNA微条形码技术,以线粒体细胞色素c氧化酶亚单位I(Cytochrome c oxidase subunit I,COI)的部分基因作为目标片段,对可食用血制品中的猪、牛、羊、马、兔、鸡、鸭、鹅、鸽共9种畜禽类动物源成分进行掺假鉴别.通过对方法的预处理及前处理、DNA提取方式、通用引物组及聚合酶链式反应(Polymerase chain reaction,PCR)扩增条件等选择与优化,经深度清洗及真空冷冻干燥处理的血制品,其基因组经提取纯化及扩增后,扩增产物经毛细管凝胶电泳确认、克隆测序,得到的序列提交至美国国家生物技术信息中心数据库进行Blast比对.对于可食用血制品中9种畜禽类动物源血成分的DNA扩增,其效率达到100％.通过这种方法对30批次市售可食用血制品进行掺假鉴定,发现其中有8批次存在其他动物源成分.该方法具有简单、可靠的特点,可作为鉴别可食用血制品中9种畜禽类动物源血成分掺假的有效检测方法.
Abstract
This study aimed to employ DNA mini-barcoding technology,using a partial mitochondrial cytochrome c oxidase subunit I(COI)gene as the target fragment,for the identification of adulterantion nine livestock and poultry blood components,including pig,cow,sheep,horse,rabbit,chicken,duck,goose,and pigeon,in edible blood products.By selecting and optimizing methods for preprocessing and pretreatment,DNA extraction protocols,universal primer sets,and polymerase chain reaction(PCR)amplification conditions,edible blood products subjected to deep cleaning and vacuum freeze-drying were processed for DNA extraction,purification,and amplification.The amplified products were confirmed through capillary gel electrophoresis and clone sequencing,and the resulting sequences were submitted to the national center for biotechnology information(NCBI)database for blast alignment.The DNA amplification efficiency for the nine livestock and poultry blood components in the edible blood products reached 100％.Using this method,adulteration detection was carried out on 30 batches of commercially available edible blood products,revealing the presence of animal-derived adulterants in 8 batches.This method is characterized by its simplicity and reliability,serving as an effective detection tool for identifying adulteration of nine livestock and poultry blood components in edible blood products.
维普
万方数据