中国媒介生物学及控制杂志 ›› 2012, Vol. 23 ›› Issue (3): 202-205.

• 论著 • 上一篇    下一篇

基孔肯雅病毒的纳米金实时荧光PCR 方法的建立

燕清丽, 杨鹏飞, 房健慧, 张丽萍, 张晓龙, 曹晓梅, 姚李四   

  1. 中国检验检疫科学研究院卫生检验检疫研究所, 北京 100123
  • 收稿日期:2011-12-31 出版日期:2012-06-20 发布日期:2012-06-20
  • 通讯作者: 姚李四,Email: yaolisi@hotmail.com
  • 基金资助:
    国家质检总局科技计划项目(2008IK200)

Development of gold nanoparticle-assisted real-time fluorescence PCR for quantitative detection of Chikungunya viruses

YAN Qing-li, YANG Peng-fei, FANG Jian-hui, ZHANG Li-ping, ZHANG Xiao-long, CAO Xiao-mei, YAO Li-si   

  1. Chinese Academy of Inspection and Quarantine, Beijing 100123, China
  • Received:2011-12-31 Online:2012-06-20 Published:2012-06-20
  • Supported by:
    Supported by the Administration of Quality Supervision Inspection and Quarantine Science and Technology Projects (No. 2008IK200)

摘要: 目的 建立一种检测基孔肯雅病毒纳米金实时荧光定量PCR的方法。方法 以课题组建立的普通实时荧光PCR方法为基础,在PCR体系中添加不同大小粒径的纳米金进行体系优化,评价优化后的体系。结果 添加纳米金的实时荧光PCR较不添加的扩增效率高;优化后的纳米金实时荧光PCR产物的Ct值与模板稀释浓度存在良好的线性关系,回归方程:y=-3.31x+41.78,R2=0.9997,PCR扩增效率为99.5%。结论 纳米金能提高实时荧光PCR的反应效率。

关键词: 纳米金, 实时荧光PCR, 基孔肯雅病毒

Abstract: Objective To develop a gold nanoparticle-assisted real-time fluorescence PCR array for quantitative detection of Chikungunya viruses. Methods On the basis of the real-time PCR that we had established, gold nanoparticles of different sizes were added to the system for optimization, which was then evaluated. Results It was found that the amplification efficacy of the system with gold nanoparticles was higher than that without gold nanoparticles, amounting to 99.5%. There existed a good linear relationship between the Ct value of the products by gold nanoparticle-assisted real-time fluorescence PCR and the concentrations of templates, with the regression equation being y=-3.31x+41.78, R2=0.9997. Conclusion Gold nanoparticle-assisted real- time fluorescence PCR has the potential to improve the efficiency of PCR amplification.

Key words: Gold nanoparticles, Real-time fluorescence PCR, Chikungunya virus

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