GI和GII诺如病毒RAA可视化快速检测方法的建立
作者:
作者单位:

(1.黑龙江八一农垦大学农学院 黑龙江大庆 163000;2.中国检验检疫科学研究院 国家市场监管重点实验室(食品质量与安全) 北京 100176;3.黑龙江八一农垦大学生命科学技术学院 黑龙江大庆 163000)

作者简介:

通讯作者:

中图分类号:

基金项目:

国家重点研发计划项目(2022YFF1100900,2022YFF0607900)


Establishment of a Visual Recombinase Aided Amplification (RAA) Method for Rapid Detection of GI and GII Noroviruses
Author:
Affiliation:

(1.College of Agriculture, Heilongjiang Bayi Agricultural University, Daqing 163000, Heilongjiang;2.Key Laboratory of Food Quality and Safety for State Market Regulation, Chinese Academy of Inspection and Quarantine, Beijing 100176;3.College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing 163000, Heilongjiang)

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:

    诺如病毒(NoV)是引起全球急性肠胃炎疾病的主要病原体之一,极易爆发传播,增加医疗与经济负担。目的:通过重组酶介导等温扩增技术(RAA)开发一种新型的NoV检测方法。方法:根据ISO TS 15216-2-2013、GB 4789.42规定的GI和GII NoV检测靶标所对应的cDNA序列,设计并筛选最佳RAA引物及探针,确定其对其它常见食源性腹泻病毒的特异性。通过优化确定最短检测时间、反应程序以及反应体系,分析该检测体系下对NoV参考质粒和真实样本检测的灵敏度,从而建立GI和GII NoV RAA可视化快速检测方法。结果:所建立的RAA检测方法特异性好,与其它食源性病毒无交叉反应。在保证扩增效率的前提下,优化后的反应程序可将检测时间缩短为10 min左右,反应成本可减少三分之二。对参考质粒和真实样本检测的灵敏度分别达10-2 ng/μL和1 ng/μL。结论:本试验建立的两种NoV检测方法特异性强、灵敏度高,且简单、快速、可视,为今后NoV快速检测奠定良好的基础。

    Abstract:

    Norovirus(NoV) is one of the major pathogens causing acute gastroenteritis diseases worldwide and is highly susceptible to outbreak transmission, increasing medical and economic burden. Objective: To develop a novel detection method for NoV by recombinase aided amplification (RAA). Methods Based on the cDNA sequences corresponding to the GI and GII NoV detection targets specified in ISO TS 15216-2-2013 and GB 4789.42, the optimal RAA primers and probes were designed and screened, and their specificity for other common food-borne diarrhea viruses was determined; the shortest detection time, reaction procedure, and reaction system were determined by optimization, and the analysis of this detection system was performed. The shortest detection time, reaction procedure and reaction system were optimized, and the sensitivity of the assay system for the detection of NoV reference plasmids and real samples was analyzed, thus establishing a rapid visualization method for the detection of GI and GII NoV RAA. The optimized reaction procedure can shorten the detection time to about 10 min and reduce the reaction cost by two-thirds, and the sensitivity of the method can reach 10-2 ng/μL for the reference plasmid and 1 ng/μL for the real sample. The two NoV assays established are specific, sensitive, simple, rapid and visualized, and provide a good basis for future rapid NoV detection.

    参考文献
    相似文献
    引证文献
引用本文

林志伟,杨艳歌,吴占文,王帅,李涛,李红娜,袁飞. GI和GII诺如病毒RAA可视化快速检测方法的建立[J].中国食品学报,2023,23(12):196-208

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:2022-12-29
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2024-01-23
  • 出版日期:
文章二维码
版权所有 :《中国食品学报》杂志社     京ICP备09084417号-4
地址 :北京市海淀区阜成路北三街8号9层      邮政编码 :100048
电话 :010-65223596 65265375      电子邮箱 :chinaspxb@vip.163.com
技术支持:北京勤云科技发展有限公司

漂浮通知


×
《中国食品学报》杂志社招聘编辑