建立及应用高通量液相芯片方法检测鉴别四种动物源性成分  

Development and Application of a High-throughput Liquid Bead Array for Simultaneous Identification of Four Animal-derived Ingredients

作　　者：梅明珠 高小博 翁文川 曹永长[3] 刘志玲 吴晓薇 段燕喻 陈茹 MEI Ming-Zhu;GAO Xiao-Bo;WENG Wen-Chuan;CAO Yong-Chang;LIU Zhi-Ling;WU Xiao-Wei;DUAN Yan-Yu;CHEN Ru(Guangzhou Customs Technology Center,Guangzhou 510623;National Research Institute for Family Planning,Beijing 100013;School of Life Science of Sun Yat-Sen University,Guangzhou 510006)

机构地区：[1]广州海关技术中心,广州510623 [2]国家卫生健康委科学技术研究所,北京100013 [3]中山大学生命科学院,广州510006

出　　处：《中国口岸科学技术》2021年第8期83-89,共7页China Port Science and Technology

基　　金：广州市科技计划(201707010487);广州海关科技项目(2018GDK04)。

摘　　要：根据GenBank中反刍动物、禽、猪和鱼线粒体DNA序列,分别设计特异性引物和探针,建立同时检测反刍动物、禽、鱼三大类物种和猪成分的四重液相芯片方法,并对其特异性、敏感性和重复试验稳定性进行验证评估。结果显示该方法能够准确检出鉴别4种目标物种,与非目标物种无交叉反应;对DNA模板检测低限可达到0.01~0.05 ng;组间检测变异系数小于20%。应用该方法检测195份饲料和肉类食品,有37份样品检出结果与标注信息不一致,经与现行标准方法比对,两者检测结果一致。研究结果表明,所建立的四重液相芯片方法特异性强、稳定性好,检测灵敏度高,能够精准检出饲料等加工产品中反刍动物、禽、猪和鱼成分。Four pairs of primers and probes against ruminant,avian,swine or fish mitochondrial DNA(mtDNA)were designed and a 4-plex liquid bead was established.The specificity,sensitivity and repeatability of this method were then investigated.The results indicated that the species of interest can be identified,differentiated and detected down to 0.01-0.05 ng of genomic DNA from specific species.The coefficients of variation(CV%)in different concentration were all less than 20%.Testing of 195 commercial samples showed a good coincidence with the method used in routine testing.The high-throughput liquid bead array established in this study,which had a high level of specificity,repeatability and sensitivity,can be employed to detect ruminant,avian,swine and fish in processed products efficiently and effectively.

关 键 词：液相芯片 反刍动物 禽 猪 鱼 动物源性成分 

分 类 号：S851.34[农业科学—预防兽医学]