摘要
根据木薯单拷贝木薯铜/锌超氧化物歧化酶(Manihotesculenta copper/zinc superoxide dismutase,mSOD2)基因序列设计特异性引物和探针,建立木薯成分微滴数字PCR定量检测方法,并测试了方法的特异性、灵敏度和适应性,并建立了拷贝数浓度(copies/μL)与木薯质量(mg)的线性关系。特异性测试表明,该方法特异于木薯成分检测,定量下限(limit of quantitation,LOQ)和检测下限(limit of detection,LOD)分别为1.67 copies/μL和0.28 copies/μL。对木薯质量百分比为5%和25%样本进行定量检测,检测结果经线性关系换算得到的木薯含量分别为5.77%和23.32%,回收率分别为115.40%和93.26%,相对标准偏差(RSD)为3.53%~4.19%。表明该方法具有良好的适用性,可对质量百分比为5%及以上木薯含量的样品进行准确定量。
In this paper,a precise quantitative droplet digital polymerase chain reaction(ddPCR)method to identify and quantify the cassava was developed.The primer pair and probe were designed based on the single copy Manihotesculenta copper/zinc superoxide dismutase(mSOD2)gene.A linear relationship between copy number concentration(copy number/μL)and the cassava mass(mg)was established.The results showed that the method was specific to the detection of cassava components.The limit of quantitation(LOQ)and limit of detection(LOD)detection limits were 1.67 copies/μL and 0.28 copies/μL respectively.According to the established linear relationship between copy number concentration(copy number/μL)and the cassava mass(mg),the quantitative analysis of 5%and 25%of the cassava mass percentage was performed,and the cassava content converted according to the linear relationship were 5.77%and 23.32%,respectively.The recovery rates were 115.40%and 93.26%,respectively,and the RSD values were between 3.53%and 4.19%.It shows that the method can accurately quantify cassava components with a mass percentage of 5%or more.It indicated that the method has good applicability in quantifying cassava.
作者
刘二龙
卢丽
魏霜
关丽军
李志勇
LIU Erlong;LU Li;WEI Shuang;GUAN Lijun;LI Zhiyong(Huangpu Customs Technology Center,Guangzhou 510730,China;Guangzhou Customs Technology Center,Guangzhou 510623,China)
出处
《食品科技》
CAS
北大核心
2020年第10期279-283,共5页
Food Science and Technology
基金
广东省科技计划项目(2017B020207008)
广州市科技计划项目(201704030125)。
关键词
木薯
微滴数字PCR
定量
物种鉴定
cassava
droplet digital PCR
species identification
quantification
作者简介
刘二龙(1978-),男,硕士研究生,研究方向为动植物分子鉴定;通讯作者:李志勇。
