流行性造血器官坏死病毒(EHNV)PCR快速检测试剂盒的研制  

Development of a PCR-based kit for detection of epizootic hematopoietic necrosis virus(EHNV)

作　　者：黄辉三[1] 李明玉[1] 母尹楠[1] 敖敬群[1] 陈新华[1] 

机构地区：[1]国家海洋局第三海洋研究所国家海洋局海洋生物遗传资源重点实验室,福建厦门361005

出　　处：《台湾海峡》2011年第1期128-131,共4页Journal of Oceanography In Taiwan Strait

基　　金：国家海洋公益性行业科研专项资助项目(200905020-03);福建省科技创新平台资助项目(2009N2003)

摘　　要：以流行性造血器官坏死病毒(EHNV)中的主衣壳蛋白基因保守区序列作为扩增靶序列,设计合成了一对特异性引物,应用快速的模板制备方法和优化PCR扩增条件,建立了EHNV病毒PCR快速检测技术,并开发成简便、快速、实用的检测试剂盒.该试剂盒的检测灵敏度相当于31个病毒粒子,模板制备时间约30 min,约4 h即可得到准确的结果;且无非特异性扩增带,适用于由EHNV病毒感染的鱼类苗种和水产品的检疫及水质环境的监测.Hematopoietic necrosis virus（EHNV） is a member of the genus Ranavirus（Iridoviridae）,which can cause visceral necrosis in a variety of larvae and adult fish,and lead to the death of fish.In this study,we designed a pair of specific primers,using a method to isolate the DNA template rapidly and we optimized the PCR amplification conditions.These primers were then used to develop a rapid and sensitive PCR based kit for the detection of EHNV virus.Using this kit,the time for preparation of the DNA template was approximately 30 minutes,and accurate results could be obtained in approximately 4 hours.The amplification bands of the PCR product were specific,and the limit of detection was approximately 31 virions.This kit is fit for rapid diagnostic tests of early infection caused by EHNV,and water quality monitoring of fishery environments.

关 键 词：生物技术 虹彩病毒 流行性造血器官坏死病毒(EHNV) PCR检测 试剂盒 

分 类 号：Q178.53[生物学—水生生物学]