应用巢式RT-PCR检测水仙退化病毒1  

Application of nested RT- PCR for detection of Narcissus degeneration virus

作　　者：金晶[1,2] 蔡伟[1] 赵爽[2] 邹文超[2] 高芳銮[2] 沈建国[1] 

机构地区：[1]福建出入境检验检疫局检验检疫技术中心,福建福州350001 [2]福建农林大学植物病毒研究所

出　　处：《植物检疫》2014年第6期59-64,共6页Plant Quarantine

基　　金：国家质检总局科技计划项目(2013IK299,2013IK094);国家级星火计划项目(2012GA720018);福建省杰出青年科学基金资助项目(2014J06008);质检公益性行业科研专项项目(201410076);福建省科技计划重点项目(2009N0001)

摘　　要：水仙退化病毒(Narcissus degeneration virus,NDV)是水仙上发生较为严重的病毒之一。本文根据NDV已报道的外壳蛋白基因序列,设计特异性引物,以感病水仙的总RNA为模板,建立了快速检测NDV的巢式RT-PCR方法。结果表明,巢式RT-PCR具有良好的特异性和灵敏度,能够从感染NDV的水仙样品上扩增出与预期大小相符的特异性目的条带,同时与其他病毒无交叉反应;灵敏度测定结果表示,巢式RT-PCR灵敏度较高,是普通RT-PCR的104倍,当RNA稀释到109倍时仍能被检测到。本文建立的巢式RT-PCR为水仙上NDV的快速检测提供了技术参考依据。Narcissus degeneration virus is one of the most serious disease of narcissus with high incidence. Two pairs of specific primers were designed and synthesized based on the published nucleotide sequence of NDV, and then the detection method of nested RT-PCR was established using the total RNA as template, which was extracted from NDV-infected narcissus sample. The results showed that nested RT-PCR was proved to have good specificity and high sensitivity. The method could amplify the target fragment from NDV-infected narcissus sample easily and successfully, no positive result was obtained from other narcissus virus samples. Sensitivity of the nested RT-PCR product indicated that it could detect NDV when total RNA was diluted to 109, higher than RT-PCR with 104 times. It suggested that nested RT-PCR which was established in this paper provided technical support for rapid detection of NDV in narcissus.

关 键 词：水仙退化病毒 检测 特异性 灵敏度 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]