中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > H5亚型禽流感病毒RT-LAMP可视化检测技术的建立 南方农业学报 2013,44 (2) 323-327
Position: Home > Articles > Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of H5 subtype  avian influenza virus Journal of Southern Agriculture 2013,44 (2) 323-327

H5亚型禽流感病毒RT-LAMP可视化检测技术的建立

作  者：
彭宜;谢芝勋;刘加波;庞耀珊;邓显文;谢志勤;谢丽基;范晴;罗思思
单  位：
广西兽医研究所
关键词：
H5亚型禽流感病毒;RT-LAMP;可视化检测;血凝素(HA)基因;荧光试剂
摘  要：
[目的]建立一种适用于H5亚型禽流感病毒(AIV)的逆转录环介导等温扩增(RT-LAMP)可视化快速检测技术,为控制疫情扩散、减少经济损失争取了时间.[方法]根据GenBank中H5亚型AIV血凝素(HA)基因序列,设计一套针对HA基因6个区域的4条特异性引物,在反应之前向反应液中加入荧光试剂(Calcein/MnCl2混合液),然后对反应条件和体系进行优化.[结果]优化的RT-LAMP技术操作简便迅速,常规水浴锅中50min可完成,能特异性地检测出H5亚型AIV,但对其他HA亚型AIV、禽呼吸道病原体无交叉扩增反应,灵敏度是常规RT-PCR的10倍;反应结束后无需打开反应管盖,即可根据反应液的颜色变化对结果进行判定.[结论]建立的H5亚型AIVRT-LAMP可视化检测技术具有简便、快速、特异等特点,可在设备有限的基层兽医站及养殖场对H5亚型AIV进行快速初步诊断.
译  名：
Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of H5 subtype  avian influenza virus
关键词：
H5 subtype avian influenza virus%RT-LAMP%visual detection%hemagglutinin(HA) gene%fluorescent reagent
摘  要：
[Objective]A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for visual detection of H5 subtype avian influenza virus (AIV) in order that epidemic could be prevented and economic loss could be reduced. [Method]According to the sequences of H5 subtype AIV hemagglutinin gene available in GenBank, four primers specific to the six sites of HA gene were designed, the fluorescent reagents (Calcein and MnCI2) were added into the reaction mixture before amplification, and the reaction conditions were optimized. [Result]The assay was performed in water bath within 50 minutes, and the sensitivity of RT-LAMP was 10-time higher than that of RT-PCR. The results showed that the assay had no cross-reaction with other HA subtype AIV and other avian respiratory pathogens. The ampli- fication result could be estimated directly by the color of reaction fluid without opening the reaction tube cap. [Conclusion] The newly developed RT-LAMP assay was simple, rapid and specific for visual detection of H5 subtype AIV in resource- limited settings like grass-roots veterinary stations and farms.

相似文章

导出引用
计量
文章访问数: 11
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46