传染性造血器官坏死病病毒RT-RAA-LFD检测方法的建立与应用  被引量:2

Development and application of an RT-RAA-LFD assay for rapiddetection of infectious haematopoietic necrosis virus

在线阅读下载全文

作  者:廖立珊 徐彬 陈兵 吴江 王津津 朱裕敏 刘荭 王婉君 孙洁 LIAO Li-shan;XU Bin;CHEN Bing;WU Jiang;WANG Jin-jin;ZHU Yu-min;LIU Hong;WANG Wan-jun;SUN Jie(Animal and Plant Inspection and Quarantine Technology Center,Shenzhen Customs,Shenzhen 518045,China;Shenzhen Institute of Quality&Safety Inspection and Research,Shenzhen 518000,China;Dalian Ocean University,Dalian 116023,China;Shenzhen Branch,China Resources WuFeng Meat Products(Henan)Co.,Ltd.,Shenzhen 518031,China)

机构地区:[1]深圳海关动植物检验检疫技术中心,广东深圳518045 [2]深圳市质量安全检验检测研究院,广东深圳518000 [3]大连海洋大学,辽宁大连116023 [4]华润五丰肉类食品(河南)有限公司深圳分公司,广东深圳518031

出  处:《中国兽医科学》2023年第8期991-996,共6页Chinese Veterinary Science

基  金:深圳市可持续发展项目(KCXFZ20211020165547010);广东省“生物安全技术”专项重点研发项目(2022B1111030001)。

摘  要:为针对传染性造血器官坏死病病毒(IHNV)建立一种可实现现场快速、准确的检测方法,结合重组酶介导扩增(RAA)和侧向流试纸条(LFD)技术,在IHNVG基因保守区域设计引物和探针,通过优化反应时间和温度等条件,建立了可用于IHNV现场可视化检测的RT-RAA-LFD方法。优化后的检测方法在35℃水浴锅中恒温反应15 min即可实现对IHNV目的基因片段的有效扩增。结果显示:该方法可以特异性地检测出IHNV,不与其他常见鱼类病毒发生交叉反应;对IHNV质粒标准品的检测下限可以达到2.15×101 copies/μL;采用该方法和实时荧光RT-PCR、实时荧光RT-RAA同时对35份临床样本进行检测,其阳性符合率为100%。上述结果表明,本研究针对IHNV成功建立了一种快速、准确、便捷的试纸条检测方法,并可适应于多种场合。To establish a rapid and accurate method for detection of infectious haematopoietic necrosis virus(IHNV),primers and probes were designed according to the conserved sequence of IHNV G gene.A real-time recombinase-aid amplification combined with lateral flow dipstick(RT-RAA-LFD)method was established for visual detection of IHNV in the field with a water bath within 15 min at 35℃after being optimized.The assay was specific with the target sequence of IHNV with a detection limit up to 2.15×101 copies/μL.Thirty-five clinical samples were tested by this method,showing 100%consistency with the real-time RT-PCR and RT-RAA.The RT-RAA-LFD assay developed in this study was a rapid,reliable and simple method which can be broadly used in the farms and diagnostic on the samples with suscepected clinical signs.

关 键 词:传染性造血器官坏死病病毒 重组酶介导扩增 侧向流试纸条 快速检测 

分 类 号:S852.659.5[农业科学—基础兽医学]

 

参考文献:

正在载入数据...

 

二级参考文献:

正在载入数据...

 

耦合文献:

正在载入数据...

 

引证文献:

正在载入数据...

 

二级引证文献:

正在载入数据...

 

同被引文献:

正在载入数据...

 

相关期刊文献:

正在载入数据...

相关的主题
相关的作者对象
相关的机构对象