基于Taqman微流控芯片技术高通量检测17种转基因玉米品系  

High-throughput Identification and Detection of 17 Transgenic Maize Events Based on Taqman Microfluidic Chip Technology

作　　者：徐君怡 曹际娟 郑秋月 杨莉莉 王永[3] 邹明强[4,5] XU Jun-Yi;CAO Ji-Juan;ZHENG Qiu-Yue;YANG Li-Li;WANG Yong;ZOU Ming-Qiang(Technology Center of Dalian Customs District,Dalian 116001,China;Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education,Dalian Minzu University,Dalian 116600,China;Tianjin Academy of Agricultural Sciences,Tianjin 300192,China;Chinese Academy of Inspection and Quarantine,Beijing 100123,China;China Inspection Laboratory Technologies Co.Ltd.,Beijing 100123,China)

机构地区：[1]大连海关技术中心,大连116001 [2]大连民族大学,生物技术与资源利用教育部重点实验室,大连116600 [3]天津市农业科学院,天津300192 [4]中国检验检疫科学研究院,北京100123 [5]中检国研(北京)科技有限公司,北京100123

出　　处：《分析化学》2020年第11期1477-1485,I0012-I0014,共12页Chinese Journal of Analytical Chemistry

基　　金：国家转基因生物新品种培育专项课题(No.2018ZX08012-001)资助。

摘　　要：将Taqman微流控芯片技术应用于实时荧光PCR平台的转基因玉米17种品系高通量检测。在一次PCR扩增过程中,以2个玉米内源基因(hmg A基因、adh 1基因)和1个上样控制基因(18S基因)为内参照,可同时完成17种转基因玉米品系(TC1507、NK603、MON87640、MON863、MON810、MIR162、GA21、DAS40278、BT176、BT11、98140、59122、3272、MON89034、MIR604、MON88017、T25)的单孔单重扩增的并行检测,检出限可达10~20 copies。本方法特异性强,灵敏度高,与“金标准”单一的实时荧光PCR方法检测结果完全吻合,具有可多样品、多靶标平行检测的优势,为转基因产品多品系混杂鉴定检测提供了高效、快速的方法。采用本方法,从16批进境的实用玉米中检出9批转基因玉米样品,且发现其中不同程度的混杂含有1~8种品系。本方法可用于口岸进境实用农产品中多品系混杂转基因的高通量检测。Taqman microfluidic chip technology was applied to the high-throughput identification and detection of 17 transgenic maize events on a real-time fluorescent PCR platform.In one PCR amplification process,2 maize endogenous genes(hmg A gene,adh 1 gene)and 1 loading control gene(18S gene)were used as internal reference,which could simultaneously complete totally 17 events of transgenic maize single-well single-plex amplification parallel detection,including TC1507,NK603,MON87640,MON863,MON810,MIR162,GA21,DAS40278,BT176,BT11,98140,59122,3272,MON89034,MIR604,MON88017 and T25,and the limit of detection reached 10-20 copies.The method showed high specificity and sensitivity,and the detection result obtained by this method was completely consistent with that of the single real-time fluorescent PCR method of"gold standard".The method had the advantages of parallel detection of multiple samples and multiple targets,and provided an efficient and rapid approach for the identification of mixture of multiple events of genetically modified products.With this method,9 batches of transgenic maize samples were detected from 16 batches of imported practical maize,containing 1-8 events.This method could be used for high-throughput detection of mixed transgenic multi-events in imported agricultural products at the port.

关 键 词：Taqman微流控芯片 转基因玉米 品系鉴定 多品系平行检测 

分 类 号：S513[农业科学—作物学]