[1]乐振窍,许泽仰,张细玲,等.FQ-PCR与IMSA检测转基因豆奶外源基因的比较研究 [J].大豆科学,2018,37(06):943-949.[doi:1011861/jissn1000-98412018060943]
　LE Zhen-qiao,XU Ze-yang,ZHANG Xi-ling,et al.Comparison of FQ-PCR and IMSA for Detecting Exogenous Genes in Genetically Modified Soymilk[J].Soybean Science,2018,37(06):943-949.[doi:1011861/jissn1000-98412018060943]
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FQ-PCR与IMSA检测转基因豆奶外源基因的比较研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第37卷 期数: 2018年06期 页码: 943-949 栏目: 出版日期: 2018-11-20
Title:
Comparison of FQ-PCR and IMSA for Detecting Exogenous Genes in Genetically Modified Soymilk
作者:
乐振窍1; 许泽仰2; 张细玲1; 陈慧国1; 祖新3
（1.福建达利食品集团有限公司，福建 惠安 362100； 2.广州迪澳生物科技有限公司，广东 广州 510507； 3.甘肃省食品检验研究院，甘肃 兰州 730030)
Author(s):
(1.Fujian Dali Food Group Co LTD, Huian 362100, China; 2.Guangzhou DIAO Bio-technology Co LTD, Guangzhou 510507, China; 3.Gansu Province Food Inspection Institute, Lanzhou 730030, China)
关键词:
FQ-PCR; IMSA; 豆奶; 外源基因; 灵敏度; 特异性
Keywords:
FQ-PCR;  IMSA;  Soymilk;  Exogenous genes;  Sensitivity;  Specificity
DOI:
1011861/jissn1000-98412018060943
文献标志码:
A
摘要:
为比较实时荧光定量PCR（FQ-PCR）与等温多自配引发扩增（IMSA）技术检测转基因豆奶中外源基因（CaMV35S、NOS、EPSPS）的灵敏度和特异性。合成CaMV35S、NOS以及EPSPS基因的质粒，通过FQ-PCR与IMSA检测定量的质粒，比较质粒灵敏度；按050%、010%、005%以及001%的比例制作转基因豆奶，通过FQ-PCR与IMSA检测转基因大豆标准品制成的豆奶，比较样品灵敏度和特异性。结果表明：IMSA检测CaMV35S、NOS、EPSPS基因的质粒灵敏度分别为3101，3101，310 cps·μL-1，FQ-PCR检测CaMV35S、NOS、EPSPS基因的质粒灵敏度皆为3101 cps·μL-1；IMSA检测CaMV35S、NOS、EPSPS基因的样品灵敏度分别为005%、005%、001%，FQ-PCR检测CaMV35S、NOS、EPSPS基因的样品灵敏度皆为010%；FQ-PCR非特异性检出DP356043的NOS以及MON87701的CaMV35S与EPSPS。说明IMSA方法检测转基因豆奶外源基因（CaMV35S、NOS、EPSPS）的灵敏度与特异性优于FQ-PCR。
Abstract:
In order to compare the sensitivity and specificity between real-time fluorescent quantitative PCR (FQ-PCR) and isothermal multiple self-matching-initiated amplification (IMSA) detecting exogenous gene (CaMV35S, NOS, EPSPS) of genetically modified soymilk The sensitivity of plasmids which were constructed with CaMV35S, NOS and EPSPS gene between FQ-PCR and IMSA were compared The genetically modified soymilk was made at a ratio of 050%, 010%, 005% and 001% to compare sample detection sensitivity and specificity between FQ-PCR and IMSA The results showed that the plasmid sensitivity of CaMV35S, NOS and EPSPS gene in genetically modified soymilk detected by IMSA were 3101, 3101, 310 cps·μL-1 respectively And the plasmid sensitivity of the three genes detected by FQ-PCR was 3101 cps·μL-1 The sample sensitivity of CaMV35S, NOS and EPSPS gene in genetically modified soymilk detected by IMSA were 005%, 005%, 001% respectively And the sample sensitivity of the three genes detected by FQ-PCR was 01% False positive of NOS in DP305423 and CaMV35S, EPSPS in MON87701 by FQ-PCR It indicated that the sensitivity and specificity of IMSA on detecting exogenous gene (CaMV35S, NOS, EPSPS) of genetically modified soymilk was much better than FQ-PCR

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备注/Memo

收稿日期：2018-08-24

基金项目：甘肃省食品药品监督管理局科研项目（2018GSFDA013）。
第一作者简介：乐振窍(1978-)，男，学士，工程师，主要从事食品安全管理研究。E-mail：zhenqiaole@daligroupcom。
通讯作者：祖新（1973-），男，学士，高级工程师，主要从事食品安全检验研究。E-mail：zuxin66@aliyuncom。

更新日期/Last Update: 2018-12-04