超分支滚环试纸检测虾传染性皮下及造血组织坏死病毒  

Detection of Infectious Hypodermal and Hematopoietic Necrosis Virus by the Hyper-Branched Rolling Circle Amplification Test Strip

作　　者：赵玉然[1] 尹伟力 岳志芹[3] 王莹[4] 李八方[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003 [2]烟台出入境检验检疫局,山东烟台264000 [3]山东出入境检验检疫局技术中心,山东青岛266002 [4]青岛农业大学食品科学与工程学院,山东青岛266109

出　　处：《武汉大学学报（理学版）》2014年第4期331-337,共7页Journal of Wuhan University:Natural Science Edition

基　　金：国家质量监督检验检疫总局科研项目(2012IK018);山东出入境检验检疫局科研项目(SK201202)

摘　　要：依据传染性皮下及造血组织坏死病毒(infectious hypodermal and hematopoietic necrosis virus,IHHNV)的非结构蛋白NS1基因序列,设计特异的锁式探针,建立IHHNV病毒超分支滚环扩增(hyper-branched rolling circle amplification,HRCA)检测方法并构建试纸条.结果表明,IHHNV HRCA试纸的检测限可以达到10拷贝/μL,较常规PCR法高约2个数量级,且能够保证对IHHNV的特异性检测.利用该试纸条对国产与进口的42份虾样本进行IHHNV检测,结果显示,HRCA试纸条在灵敏度上优于常规PCR,且方法直观、更易于结果判定.Based on the nonstructural proten NS1 gene sequence of infectious hypodermal and hematopoietic nec- rosis virus （IHHNV）, a specific padlock probe was designed, and the hyper-branched rolling circle amplification （HR- CA） assay and the test strip for IHHNV were established. The results show that the detection limit of the present HRCA test strip is close to 10 copies//~L which is two orders of magnitude higher than conventional polymerase chain reaction （PCR）, and specific to IHHNV. In practical applications, 42 patches of shrimps were collected to detect IH- HNV at home and abroad by the present test strip, and the result showed that the HRCA test strip was more sensi- tive, intuitive and easy to analysize than conventional PCR.

关 键 词：传染性皮下及造血器官坏死病毒 锁式探针 超分支滚环扩增 检测试纸 

分 类 号：S945.4[农业科学—水产养殖]