超分支滚环试纸检测虾传染性皮下及造血组织坏死病毒

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 赵玉然 ZHAO Yuran 1 first-author 尹伟力 YIN Weili 2 岳志芹 YUE Zhiqin 3 王莹 WANG Ying 4 李八方 LI Bafang 1 中国海洋大学食品科学与工程学院 中国海洋大学食品科学与工程学院 College of Food Science and Engineering,Ocean University of China College of Food Science and Engineering,Ocean University of China 烟台出入境检验检疫局 烟台出入境检验检疫局 Yantai Entry-Exit Inspection and Quarantine Bureau Yantai Entry-Exit Inspection and Quarantine Bureau 山东出入境检验检疫局技术中心 山东出入境检验检疫局技术中心 Technical Centre of Shandong Entry-Exit Inspection and Quarantine Bureau Technical Centre of Shandong Entry-Exit Inspection and Quarantine Bureau 青岛农业大学食品科学与工程学院 青岛农业大学食品科学与工程学院 College of Food Science and Engineering,Qingdao Agriculture University College of Food Science and Engineering,Qingdao Agriculture University 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 protein NS1gene sequence of infectious hypodermal and hematopoietic necrosis virus（IHHNV）,a specific padlock probe was designed,and the hyper-branched rolling circle amplification（HRCA）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 10copies/μL which is two orders of magnitude higher than conventional polymerase chain reaction（PCR）,and specific to IHHNV.In practical applications,42patches of shrimps were collected to detect IHHNV at home and abroad by the present test strip,and the result showed that the HRCA test strip was more sensitive,intuitive and easy to analysize than conventional PCR. 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 IHHNV（infectious hypodermal and hematopoietic necrosis virus） padlock probe HRCA（hyper-branched rolling circle amplification） test strip 10.14188/j.1671-8836.2014.04.023 S945.4 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） 2014-04-01 2021-04-01 4 60