多重PCR-DHPLC法检测转基因棉花品系  

Simultaneously detection of three genetically modified cotton events by Multiplex PCR Coupled with Denaturing High Performance Liquid Chromatography(mPCR-DHPLC)

作　　者：向才玉[1,2] 凌杏园[1] 潘广[1] 康林[1,2] 陈冬美[1] 李秋枫[1] 刘新娇[1] 欧阳辉 章桂明[1,2] 

机构地区：[1]深圳出入境检验检疫局,广东深圳518045 [2]深圳市检验检疫科学研究院深圳市外来有害生物检测技术研发重点实验室 [3]深圳博睿祥晖生物技术有限公司

出　　处：《植物检疫》2015年第1期37-41,共5页Plant Quarantine

基　　金：国家"十二五"转基因重大专项(2013ZX08012-001-002-003)

摘　　要：根据3种转基因棉花品系的边界序列设计品系检测引物,建立了一种特异性检测转基因棉花品系MON531、MON1445和MON15985的多重PCR-DHPLC方法。以20种不同的转基因及非转基因作物DNA验证该方法的特异性,结果只有MON531、MON15985和MON1445有特异的品系扩增片段峰,而其他转基因和非转基因作物无品系扩增片段峰,表明该方法特异性强。灵敏度实验结果表明3种转基因棉花的检测下限均为1 ng,灵敏度高。建立的方法可用于转基因棉花MON531、MON1445和MON15985品系及含有其成分产品的筛查或定性检测。To discriminate the GM cotton events efficiently, an analysis method combining multiplex PCR and DHPLC for simultaneously identifying GM cotton events, MON531, MON1445 and MON15985, was developed by designing the PCR primer sets on the boundary DNA sequences of the three events. The specifi city of this method was tested by using twenty kinds of different GM and non-GM crops. The result showed that only MON531, MON1445 and MON15985 could obtain eluting peaks that are corresponding to the expected size, and other GM and nonGM materials obtained no PCR product, indicating, the specifi city of this multiplex detection method was high. Thesensitivity test results showed that the eluting peaks could be observed when 1 ng more genomic DNA of the three GMO events was used as the PCR templates, showing the limit of this detection method is 1 ng. The method is a reliable and promising tool for simultaneously screening and identifi cation of the three GM cotton events from the agricultural products and other processed products.

关 键 词：转基因棉花 多重PCR DHPLC 检测 

分 类 号：S562[农业科学—作物学]