水产科学
水產科學
수산과학
FISHERIES SCIENCE
2014年
3期
157-162
,共6页
尹伟力%耿金培%刘宁%岳志芹%郑小龙%刘荭%许红岩
尹偉力%耿金培%劉寧%嶽誌芹%鄭小龍%劉葒%許紅巖
윤위력%경금배%류저%악지근%정소룡%류홍%허홍암
真鲷虹彩病毒%锦鲤疱疹病毒%液相芯片%检测
真鯛虹綵病毒%錦鯉皰疹病毒%液相芯片%檢測
진조홍채병독%금리포진병독%액상심편%검측
Red Sea Bream Iridovirus(RSIV)%Koi Herpesviru(KHV)%liquidchip%detection
为建立一种同时检测真鲷虹彩病毒、锦鲤疱疹病毒的液相芯片快速检测技术,用DNAStar软件对GenBank中真鲷虹彩病毒的CP基因和锦鲤疱疹病毒的TK基因进行序列分析,设计真鲷虹彩病毒、锦鲤疱疹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真鲷虹彩病毒、锦鲤疱疹病毒PCR产物杂交反应,用液相芯片仪器检测荧光信号。试验结果表明,液相芯片检测体系能够正确的检测出真鲷虹彩病毒、锦鲤疱疹病毒。真鲷虹彩病毒、锦鲤疱疹病毒病毒核酸的最低检出量为10 pg和100 pg ,检测特异性高,说明初步建立了同时检测真鲷虹彩病毒、锦鲤疱疹病毒的液相芯片技术,为进一步构建其他水生动物病原体快速高通量检测平台奠定基础。
為建立一種同時檢測真鯛虹綵病毒、錦鯉皰疹病毒的液相芯片快速檢測技術,用DNAStar軟件對GenBank中真鯛虹綵病毒的CP基因和錦鯉皰疹病毒的TK基因進行序列分析,設計真鯛虹綵病毒、錦鯉皰疹病毒特異性引物併標記生物素。探針氨基化脩飾,與熒光編碼微毬偶聯後與真鯛虹綵病毒、錦鯉皰疹病毒PCR產物雜交反應,用液相芯片儀器檢測熒光信號。試驗結果錶明,液相芯片檢測體繫能夠正確的檢測齣真鯛虹綵病毒、錦鯉皰疹病毒。真鯛虹綵病毒、錦鯉皰疹病毒病毒覈痠的最低檢齣量為10 pg和100 pg ,檢測特異性高,說明初步建立瞭同時檢測真鯛虹綵病毒、錦鯉皰疹病毒的液相芯片技術,為進一步構建其他水生動物病原體快速高通量檢測平檯奠定基礎。
위건립일충동시검측진조홍채병독、금리포진병독적액상심편쾌속검측기술,용DNAStar연건대GenBank중진조홍채병독적CP기인화금리포진병독적TK기인진행서렬분석,설계진조홍채병독、금리포진병독특이성인물병표기생물소。탐침안기화수식,여형광편마미구우련후여진조홍채병독、금리포진병독PCR산물잡교반응,용액상심편의기검측형광신호。시험결과표명,액상심편검측체계능구정학적검측출진조홍채병독、금리포진병독。진조홍채병독、금리포진병독병독핵산적최저검출량위10 pg화100 pg ,검측특이성고,설명초보건립료동시검측진조홍채병독、금리포진병독적액상심편기술,위진일보구건기타수생동물병원체쾌속고통량검측평태전정기출。
The CP gene of Red Sea Bream Iridovirus (RSIV) and TK gene of Koi Herpesvirus (KHV) in the GenBank was analysed using the software DNAStar 7 .0 ,and specific RSIV and KHV primers labeled with biotin was prepared and coupled with fluorescence-coded microspheres to develop liquidchip technique to detect RSIV and KHV simultaneously .The aminationed probe was used for hybridization reaction to PCR products of RSIV and KHV ,then Liquidchip detection technique for detection of RSIV and KHV was established by BD FACSArray to detect fluorescence signal in the reaction system .The results showed that RSIV and KHV was accurately and high specifically detected by liquidchip detection technique with minimal limit of 10 pg of RSIV DNA and 100 pg of KHV DNA .The Liquidchip detection technique for detection of RSIV ,and KHV was established preliminarily ,which provided a foundation for further research on rapidly high throughout detection of other aquatic animal pathogens .