[1]陈宏备,施开创,李向涛,等.小鼠IL-1β、TNF-α TaqMan荧光定量RT-PCR检测方法的建立及脑心肌炎病毒感染小鼠的检测[J].中国预防兽医学报,2011,(07):531-536.
 CHEN Hong-bei,SHI Kai-chuang,LI Xiang-tao,et al.Development of a TaqMan real-time PCR assay for detection of IL-1β and TNF-α mRNA in mice experimentally infected with encephalomyocarditis virus[J].Chinese journal of preventive veterinary medicine,2011,(07):531-536.
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小鼠IL-1β、TNF-α TaqMan荧光定量RT-PCR检测方法的建立及脑心肌炎病毒感染小鼠的检测
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年07期
页码:
531-536
栏目:
诊断和检测技术
出版日期:
2011-07-15

文章信息/Info

Title:
Development of a TaqMan real-time PCR assay for detection of IL-1β and TNF-α mRNA in mice experimentally infected with encephalomyocarditis virus
作者:
陈宏备;施开创;李向涛;郑敏;郑喜邦;李军;
广西大学动物科学技术学院;广西动物疫病预防控制中心;
Author(s):
CHEN Hong-bei1 SHI Kai-chuang2 LI Xiang-tao1 ZHENG Min2 ZHENG Xi-bang1 LI Jun2
1. College of Animal Science and Technology, Guangxi University, Nanning 530005, China; 2. Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China
关键词:
脑心肌炎病毒小鼠促炎细胞因子荧光定量RT-PCR
Keywords:
encephalomyocarditis virus mouse proinflammatory cytokine real-time RT-PCR
摘要:
为探讨脑心肌炎病毒(EMCV)感染后促炎细胞因子的表达水平、从分子水平深入研究EMCV的致病机制,本研究分别建立了检测小鼠IL-1β、TNF-α和管家基因β-actin 的 TaqMan real-time PCR 检测方法。该方法标准曲线的相关系数均达到0.998以上,检出下限均达到10copies/μL质粒标准品,组内与组间的变异系数均小于2%。应用该方法对猪源 EMCV GXLC 株人工感染小鼠的脑、心、脾中IL-1β、TNF-α mRNA 的转录水平进行检测,发现感染后第4dIL-1β、TNF-α mRNA 的转录水平达到峰值,并且与小鼠发病死亡高峰存在明显的时间相关性。本研究所建立的TaqMan real-time PCR 检测方法为小鼠促炎细胞因子的检测及定量分析提供了技术手段。
Abstract:
In this study, a real-time RT-PCR assay based on TaqMan probe for detection of mouse proinflammatory cytokine gene IL-1β and TNF-α was established, respectively. The assays were highly specific, sensitive and reproducible, of which the correlation coefficient of the standard curve was over 0.998, the sensitivity was 10 copies/μL of standard recombinant plasmid and the coefficient of variation was less than 2 percent for both intra-assay and inter-assay. The established assays were used to detect IL-1β and TNF-α mRNA levels in brain, heart and spleen tissues of mice experimentally infected with porcine encephalomyocarditis virus (EMCV) GXLC strain. The results showed that IL-1β and TNF-α mRNA expression levels reached peak value at 4 day post EMCV infection, with a time correlation between the expression levels and the mortality of infected mice. The results indicated that the TaqMan real-time PCR assay could be used as an effective tool for detection and quantification of these proinflammatory cytokines

参考文献/References:

[1] 施开创,屈素洁,陈进喜,许瑞胜,郑敏,刘棋,陈汉忠,李刚. 猪源脑心肌炎病毒GXLC株全基因组序列测定与分析[J]. 病毒学报. 2010年02期
[2] 盖新娜,杨汉春,郭鑫,吕艳丽,王芳,陈艳红,查振林. 猪脑心肌炎病毒的分离与鉴定[J]. 畜牧兽医学报. 2007年01期

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备注/Memo

备注/Memo:
基金:广西科学基金项目(桂科青0728047);;广西科技创新能力与条件建设基金项目(08-05-01D)
更新日期/Last Update: 2011-09-29