加工肉制品自动化DNA提取体系建立及其验证  

Establishment and Validation of An Automated DNA Extraction System for Processed Meat Products

作　　者：姜展樾 乔倩[1] 陈鹏 马丽侠[1] 蒋子敬 叶德萍[1] 周李华[1] 韩国全[2] JIANG Zhanyue;QIAO Qian;CHEN Peng;MA Lixia;JIANG Zijing;YE Deping;ZHOU Lihua;HAN Guoquan(National Institute of Measurement and Testing Technology,Chengdu 610021,China;College of Food,Sichuan Agriculture University,Ya’an 625000,China)

机构地区：[1]中国测试技术研究院,四川成都610021 [2]四川农业大学食品学院,四川雅安625000

出　　处：《食品科技》2021年第8期104-110,共7页Food Science and Technology

基　　金：国家重点研发计划项目(2018YFC1603400)。

摘　　要：针对加工肉制品中DNA提取难度大、干扰杂质多、难以实现自动化、不能满足高通量的核酸检测需求等问题,研究建立了一种适用于加工肉制品的高通量、自动化的DNA提取方法。该方法采用实验室设计优化磁珠并对体系关键组分进行优化,考察了方法适用性和重复性,与国内外典型厂商的5种试剂盒比对,再联合8家实验室验证了其对39种加工肉制品的提取效果。结果表明,此方法提取的加工肉制品DNA浓度高、纯度好,适用于动物组织及加工肉制品手动和自动化提取,实际样品检出率为100%。该方法重复性好,适用于不同种类动物组织及加工肉制品自动化、高通量的DNA提取,产物满足荧光定量PCR检测、二代测序等下游试验要求,为食品掺假的分子鉴别提供技术支撑。Processed meat products have many interfering impurities,which is difficult to achieve automation and high throughput nucleic acid detection requirements.This study established a high-throughput,automated DNA extraction method for processed meat products.In this method,magnetic beads were optimized by laboratory design and key components of the system were optimized.The applicability and reproducibility of the method were investigated.Compared with 5 kits from typical manufacturers at home and abroad,the extraction effects of 39 kinds of processed meat products were verified by 8 laboratories.The results showed that this method was suitable for manual and automatic extraction of animal tissues and processed meat products with high concentration and good purity.The actual sample detection rate was 100%.This method has good repeatability and is suitable for automatic and high-throughput DNA extraction from different animal tissues and processed meat products.The product meets the downstream test requirements such as fluorescence quantitative PCR detection and second-generation sequencing,providing technical support for molecular identification of food adulteration.

关 键 词：磁珠法 DNA提取 实时荧光PCR 加工肉制品 自动化 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]