CAJ | 학술논문

为建立一种同时检测对虾桃拉病毒、虾黄头病毒的液相芯片快速检测技术，用DNAStar软件对GenBank中对虾桃拉病毒的CP2基因和虾黄头病毒的N基因进行序列分析，设计对虾桃拉病毒、虾黄头病毒特异性引物并标记生物素。探针氨基化修饰，与荧光编码微球偶联后与对虾桃拉病毒、虾黄头病毒病毒RT-PCR产物杂交反应，用液相芯片仪器检测荧光信号。该检测体系对对虾桃拉病毒、虾黄头病毒病毒核酸检测灵敏度高，其最低检测限为100 pg ，且与白斑病毒、鲤春病毒血症病毒、传染性造血器官坏死病毒核酸等无交叉反应。该方法灵敏度及特异性高，为同时快速检测对虾桃拉病毒、虾黄头病毒提供了简捷快速的技术手段。
위건립일충동시검측대하도랍병독、하황두병독적액상심편쾌속검측기술，용DNAStar연건대GenBank중대하도랍병독적CP2기인화하황두병독적N기인진행서렬분석，설계대하도랍병독、하황두병독특이성인물병표기생물소。탐침안기화수식，여형광편마미구우련후여대하도랍병독、하황두병독병독RT-PCR산물잡교반응，용액상심편의기검측형광신호。해검측체계대대하도랍병독、하황두병독병독핵산검측령민도고，기최저검측한위100 pg ，차여백반병독、리춘병독혈증병독、전염성조혈기관배사병독핵산등무교차반응。해방법령민도급특이성고，위동시쾌속검측대하도랍병독、하황두병독제공료간첩쾌속적기술수단。
The CP2 gene of Taura syndrome virus (TSV) and N gene of yellow head disease virus(YHDV) in the GenBank were analysed by the software DNAStar 7 .0 ,and specific TSV and YHDV primers labeled with biotin were prepared and coupled with fluorescence-coded microspheres to develop liquichip technique to detect TSV and YHDV simultaneously .The aminationed probe was used for hybridization reaction to PCR products of TSV and YHDV ,and then the liquichip detection technique for detection of TSV and YHDV was established using BD FACSArray to detect fluorescence signal in the reaction system .This assay was highly sensitive to TSV ,and YHDV with a lower detection limit of 100 pg ,and was not susceptible to cross reaction with other viruses ,including white spot syndrome virus (WSSV ) ,spring viremia of carp virus (SVCV ) ,and infectious haematopoietic necrosis virus (IHNV ) .In conclusion ,the liquichip technique assay is sensitive ,and specific for TSV and YHDV detection ,and provides a novel , convenient ,and rapid approach for the detection of TSV and YHDV .