多重real-time PCR技术快速鉴别特种乳中的乳源动物成分  

Multiple Real-time Polymerase Chain Reaction for Identification of Animal-derived Ingredients in Milk Products from Minor Dairy Species

作　　者：杨艳歌[1] 李莉[1] 王丹丹 李唯熙 王洪越 刘鸣畅[1] 吴亚君[1] YANG Yange;LI Li;WANG Dandan;LI Weixi;WANG Hongyue;LIU Mingchang;WU Yajun(Chinese Academy of Inspection and Quarantine,Beijing 100176,China;College of Life Sciences and Food Engineering,Hebei University of Engineering,Handan 056038,China)

机构地区：[1]中国检验检疫科学研究院,北京100176 [2]河北工程大学生命科学与食品工程学院,河北邯郸056038

出　　处：《食品科学》2021年第16期312-321,共10页Food Science

基　　金：中国检验检疫科学研究院基本科研业务费项目(2018JK027);“十三五”国家重点研发计划重点专项(2016YFD0401203)。

摘　　要：建立一种特种乳中乳源动物成分快速鉴别多重实时聚合酶链式反应技术。通过筛选建立8种不同乳源物种检测的多重实时聚合酶链式反应方法,在一个反应体系里可同时检测4种乳源的特异性靶基因,绝对灵敏度达0.1~5 pg/μL,检出限可达0.1%。同时,建立了高效的DNA提取方法,样品裂解后可在12 min左右完成96个样品的DNA提取,大大缩短了样品前处理时间。采用该方法对市售特色乳产品进行调查,结果显示标识不准确的产品比例达36.36%。The aim of this study was to establish a multiple real-time polymerase chain reaction(real-time PCR)method for the rapid identification of animal-derived ingredients in milk products from minor dairy species.A multiple real-time PCR system was developed for eight dairy animal-derived ingredients,which allowed for the simultaneous detection of four target genes.The absolute sensitivity was 0.1-5 pg/μL and the limit of detection(LOD)was 0.1%.Besides,an efficient DNA extraction method for dairy products was established,and DNA extraction from 96 samples could be completed within 12 min after sample lysis,greatly shortening the sample pretreatment time.By using this method,we surveyed commercially available dairy products from minor dairy species,revealing that 36.36%of the samples were inaccurately labelled.

关 键 词：多重实时聚合酶链式反应 特种乳 乳源动物 掺伪 鉴别 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]