四种基因定量方法对实时荧光PCR与微滴式数字PCR检测霍乱弧菌基因表达量的分析

被引:6
作者
唐静 [1 ]
贾俊涛 [1 ]
赵丽青 [1 ]
王静 [2 ]
张健 [1 ]
姜英辉 [1 ]
徐彪 [1 ]
王昌军 [1 ]
马云 [1 ]
机构
[1] 山东出入境检验检疫局检验检疫技术中心食品农产品检测中心
[2] 威海出入境检验检疫局
关键词
基因表达; 实时荧光定量PCR; 微滴式数字PCR;
D O I
10.13982/j.mfst.1673-9078.2017.3.015
中图分类号
R440 [];
学科分类号
100208 ;
摘要
基因表达量测定对基因功能解析和探索生命机理具有重大意义,目前实时荧光PCR和微滴式数字PCR技术是进行基因表达量测定的有效方法。本文以低温处理的霍乱弧菌sfs、vcc、Rec A、hly及16S r RNA基因为研究对象,分别用实时荧光PCR和微滴式数字PCR技术对低温处理前后霍乱弧菌的上述基因表达量进行测定,实时荧光PCR实验数据分别采用??CT和ge Norm法进行分析,微滴式数字PCR实验数据分别采用相对定量和绝对定量法进行分析,获得处理组基因表达量相对于对照组的变化倍数;使用多维尺度法来分析四种基因定量方法的数据。结果表明,在本实验条件下,四种基因定量方法的分析结果存在差异;16S r RNA基因表达量发生了变化,不适合作为内参基因;微滴式数字PCR能更直观的给出基因表达量变化的结果,并且基因表达差异相对定量分析的准确度更高。
引用
收藏
页码:93 / 98+80 +80
页数:7
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