本平台为互联网非涉密平台,严禁处理、传输国家秘密、工作秘密或敏感信息

GⅡ型诺如病毒微滴式数字RT-PCR检测方法的建立及初步应用
CSTR:
作者:
作者单位:

作者简介:

管锦绣(1993-),女,硕士研究生在读,主要研究方向为食品微生物 通讯作者:翁文川(1973-),男,高级工程师,主要研究方向为食品与化妆品质量安全及微生物检测

通讯作者:

中图分类号:

基金项目:

广州市科信委项目(2014J4500027)


Establishment and Preliminary Application of Droplet Digital RT-PCR Assay for Detection of GⅡ Norovirus
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    为了建立快速准确的GⅡ型诺如病毒定量检测方法,本研究将微滴式数字RT-PCR技术应用于GⅡ型诺如病毒检测中,并与实时荧光RT-PCR法进行了比对。微滴式数字RT-PCR反应退火温度的优化实验确定了其最佳退火温度为56 ℃,实时荧光RT-PCR和微滴式数字RT-PCR两种方法的灵敏度实验对比表明微滴式数字RT-PCR法检测GⅡ型诺如病毒的灵敏度为5.40 copies/μL,其灵敏度高于实时荧光RT-PCR法,重复性实验对比表明两种方法在检测中间浓度的GⅡ型诺如病毒时重复性均较好;人工污染西生菜实验中表明微滴式数字RT-PCR法最低检测限为54.00 copies/μL。本研究建立的GⅡ型诺如病毒的微滴式RT-PCR检测方法,灵敏度高,重复性良好,在人工污染西生菜GⅡ型诺如病毒的检测中表现理想,具有良好的应用前景。

    Abstract:

    The droplet digital RT-PCR assay was applied in the detection of GⅡnorovirus to establish a rapid and accurate method in this study, which was compared with real-time fluorescent RT-PCR. The annealing temperature of droplet digital RT-PCR was optimized and determined to be 56℃. Compared with real-time fluorescent RT-PCR, the sensitivity of droplet digital RT-PCR was determined to be 5.40 copies/μL which was higher than that of real-time fluorescent RT-PCR, and the repeatability of real-time fluorescent RT-PCR and droplet digital RT-PCR were good determined by the comparison test. The droplet digital RT-PCR was applied to detect the artificially contaminated Romaine Lettuce with the sensitivity wof 54.00 copies/μL. Droplet digital RT-PCR assay established for detection of GⅡnorovirus had a high sensitivity and repeatability, and had an outstanding performance in the detection of the artificially contaminated Romaine Lettuce, which would own a good application prospects.

    参考文献
    相似文献
    引证文献
引用本文

管锦绣,翁文川,许喜林,凌莉,冼钰茵,易蓉,李志勇. GⅡ型诺如病毒微滴式数字RT-PCR检测方法的建立及初步应用[J].现代食品科技,2018,34(5):222-228.

复制
分享
相关视频

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:2017-11-18
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2018-06-04
  • 出版日期:
文章二维码