+高级检索
首页 > 过刊浏览>2020年第30卷第3期 >25-31. DOI:10. 3969 / j.issn.1671-7856. 2020. 03. 005
PDF HTML阅读 XML下载 导出引用 引用提醒
实时荧光定量 RT-PCR 方法在登革热小鼠模型中的评估及应用
作者:
作者单位:

北京协和医学院比较医学中心,中国医学科学院医学实验动物研究所,新发再发传染病动物模型研究北京市重点 实验室,卫健委人类疾病比较医学重点实验室,北京 100021

中图分类号:

R-33


Evaluation of a real-time fluorescence quantitative PCR in the dengue mouse model
Author:
  • CHEN Qian

    CHEN Qian

    Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • LI Dan

    LI Dan

    Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • TONG Wei

    TONG Wei

    Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • CAI Fangzhou

    CAI Fangzhou

    Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • GUO Zhi

    GUO Zhi

    Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • BAO Linlin

    BAO Linlin

    Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • WANG Wei

    WANG Wei

    Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China
    在期刊界中查找
    在百度中查找
    在本站中查找
Affiliation:

Comparative Medicine Center, Peking Union Medical College (PUMC). Institute of Medical Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS). Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious. NHC Key Laboratory of Human Disease Comparative Medicine, Beijing 100021, China

  • 摘要
    • | |
  • 访问统计
    • |
  • 参考文献
    • |
  • 相似文献
    • |
  • 引证文献
    • | |
  • 文章评论
    摘要:

    目的 评估登革病毒实时荧光定量 RT-PCR 方法,应用于登革热小鼠模型关键指标的检测。 方法 首先,选择合适的特异引物和探针,分子生物方法制备质粒标准品,优化 PCR 体系和反应条件;其次,评估该实时荧光定量 RT-PCR 方法的灵敏性、特异性及稳定性;最后,验证该方法对登革病毒感染小鼠血清和组织样本的适用性。 结果 成功确定一步法实时荧光定量 RT-PCR 方法;该方法较为灵敏,最低检测线为 49. 6 Copies/ μL;方法特异性好,未见非特异性扩增;方法稳定性好,样本 Ct 值标准差均小于 0. 5,且 CV ﹤ 5%;登革病毒感染小鼠模型样品的检测结果符合实验预期。 结论 该 qRT-PCR 方法可作为关键指标检测方法,用于登革热小鼠模型病毒的定量检测及评估。

    Abstract:

    Objective To evaluate a real-time fluorescence quantitative PCR of dengue virus by detection of key indicators in the dengue mouse model. Methods Appropriate specific primers and probes were selected, the molecular method was used to prepare the plasmid standard, and the PCR system and reaction conditions were optimized. Then, the sensitivity, specificity, and stability of the real-time fluorescence quantitative PCR method were evaluated. Finally, this method were tested by using serum and tissue samples from dengue virus-infected mice. Results One-step real-time fluorescence quantitative PCR was performed successfully. The method was sensitive, and the lowest detection limit was 49. 6 copies/ μL. The method was specific, and non-specific amplification was not observed. The method repeatability was good, the standard deviation of sample Ct values was less than 0. 5, and the CV was <5%. The test result of samples from the dengue virus-infected mice were in line with experimental expectations. Conclusions The qRT-PCR method can be used for quantitative detection and evaluation of dengue virus in a mouse model.

    参考文献
    相似文献
    引证文献
引用本文

陈 倩,李 丹,佟 巍,蔡方舟,郭 智,鲍琳琳,王 卫.实时荧光定量 RT-PCR 方法在登革热小鼠模型中的评估及应用[J].中国比较医学杂志,2020,30(3):25~31.

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2019-10-08
  • 在线发布日期: 2020-08-19
您是第 位访问者
版权所有:中国比较医学杂志 主管单位：中国科学技术协会 主办单位：中国实验动物学会 中国医学科学院医学实验动物研究所
地址：北京市朝阳区潘家园南里5号 邮 编 ：100021 电话：010-67779337 E-mail:bjb@cnilas.org
技术支持：北京勤云科技发展有限公司
防诈骗提示！请勿点击不明链接或添加个人微信。编辑部所有邮箱后缀均为@cnilas.org
关闭