PCR方法制备地高辛标记DNA探针检测中国对虾非包涵体型杆状病毒

doi:10.13242/j.cnki.bingduxuebao.001211

IOCAS-IR

	PCR方法制备地高辛标记DNA探针检测中国对虾非包涵体型杆状病毒
	徐洪涛,朴春爱,杨朵,王雷,张新宇,侯云德
发表期刊	病毒学报
ISSN	11-3424/R
	2000-03-30
出版年	2000
期号	01 页码:77-79
文献类型	CNKI期刊论文
摘要	A pair of primers created from information of PmNOBⅢ genome DNA Sal I fragment produced a 355bp band by using Penaeus chinensis non occluded baculovirus (PcNOBV),the WSBV isolate from P.hinensis in mainland China,as the DNA template.The specific PCR product was cloned,sequenced and labeled with digoxigenin (DIG)DNA labeling kit(Boehringer Mannheim).The DIG labeled fragment was tested by dot blot hybridization for sensitivity and specificity with purified PcNOBV nucleocapsid,PcNOBV infected shrimp tissues and healthy shrimp tissues.The detection limit of the DNA probe is 6.8pg of purified PcNOBV DNA.No hybridization signals were observed using DNA from healthy shrimp as template.Healthy P.chinensis,artificially infected P.chinensis and pond reared adult P.chinensis were screened for PcNOBV infection by both PCR and the hybridization assay.The results showed a good relationship between PCR and the hybridization assay.These findings demonstrate that the DIG labeled probe can be used as a sensitive,specific and cost effective reagent for detection of PcNOBV.
关键词	中国对虾中国对虾非包涵体型杆状病毒(Pc NOBV) DNA探针斑点杂交聚合酶链反应(PCR)
CNKI专辑号	A;E;D
CNKI专辑名称	基础科学;医药卫生科技;农业科技
CNKI专题号	D052;
CNKI专题名称	水产和渔业;
分类号	S945.19
收录类别	北大核心
语种	中文;
DOI	10.13242/j.cnki.bingduxuebao.001211
资助项目(基金）	海洋 8 63 ( 819-0 2 -0 5 ) ; 病毒基因工程国家重点实验室客座课题
引用统计
文献类型	CNKI期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/197587
专题	中国科学院海洋研究所
作者单位	1.中国科学院海洋研究所!山东青岛266071 2.中国预防医学科学院病毒学研究所!北京100052 3.西安医科大学!陕西西安710061
推荐引用方式 GB/T 7714	徐洪涛,朴春爱,杨朵,王雷,张新宇,侯云德. PCR方法制备地高辛标记DNA探针检测中国对虾非包涵体型杆状病毒. 2000.