高通量二代测序基因条形码技术在动物源性制品物种鉴定中的应用

Application of High-throughput Sequencing and Gene Barcode Technology in Species Identification of Animal-derived Products

该研究利用二代测序和DNA条形码技术对生物制品中动物源性成分鉴定方法进行探索性研究。首先对常见的哺乳动物、禽类、鱼类物种以及多种混合样本进行了核酸提取,并利用PCR技术对其线粒体基因16S rRNA区域354 bp的检测位点进行扩增。使用Illumina Miseq二代测序技术获取所有序列信息,并与Gen Bank数据库比对分析样本中的物种组成。结果显示,混合样品中所有动物源性成分均得到正确鉴定;鹅和鸭核酸的最低检测下限为0.5ng/μL;市售商业化动物源性制品经检测发现2起标签不符问题。该检测方法在一个高通量测序和结果比对分析实验周期内,实现了混合DNA片段序列信息的一次性获取。检测结果准确,适用范围广,有望为动物制品标识符合性检查、打击走私等方面提供技术保障。

This study used high-throughput sequencing and DNA barcoding technology to make an exploratory study on the identification method of animal-derived components in biological products. Nucleic acid was extracted from common mammals, avian, fish species and a variety of mixed samples. PCR technology was used to amplify the 354 bp detection site of the mitochondrial gene 16 S rRNA region. Illumina Miseq sequencing technology was used to obtain all sequence information, and the species composition in the samples was compared with Gen Bank database. The results showed that all the components of animal origin in the mixed samples were correctly identified;the minimum detection limit of nucleic acid of goose and duck was 0.5 ng/μL;two labels of commercial products of animal origin were found to be inconsistent after testing. The detection method achieved a one-time acquisition of mixed DNA fragment sequence information in a high-throughput sequencing and result comparison analysis experimental cycle. The test results are accurate and applicable in a wide range, and it is expected to provide technical support for animal product identification compliance inspection and anti-smuggling.