生物素肼化学标记和DIG标记cDNA探针检测柑桔裂皮病类病毒  

BIOTIN HYDRAZIDE-LABELLED AND DIG-LABELLED cDNA PROBES FOR THE DIAGNOSIS OF CITRUS EXCORTIS VIROID IN PLANTS

作　　者：胡勤学[1,2,3] 张春立[1,2,3] 陈捷 吴德喜 林木兰[1,2,3] 

机构地区：[1]中国科学院武汉病毒研究所 [2]武汉同济医大附属协和医院 [3]湖南省园艺所

出　　处：《病毒学报》1997年第2期159-163,共5页Chinese Journal of Virology

基　　金：国家"八五"攻关项目

摘　　要：分别用生物素肼化学标记和ＤＩＧ标记我国柑桔裂皮病类病毒（ＣＥＶｄ）全长克隆ｃＤＮＡ，用以上探针对不同来源的核酸样品进行斑点杂交。两种探针可检出病柑桔总核酸的最低含量约为４００ｎｇ／斑点和８０ｎｇ／斑点；生物素肼标记ＣＥＶｄ－ｃＤＮＡ探针，可检出ＣＥＶｄ－ｃＤＮＡ的最低含量约为１０ｐｇ／斑点。研制的ＣＥＶｄ检测试剂盒能检测出发病和隐性带毒苗木中的ＣＥＶｄ，灵敏、特异、简便、快速、试剂盒已被用于检测来自我国主要裂皮病病区、湖南、湖北、福建等地的柑桔和香橼材料。Biotin hydrazide chemical labelling and DIG labelling were used to prepare nonradioactive nucleic acid probes respectively.The cloned full-length CEVd-cDNA isolated from China was labelled.Using biotin hydrazide-labelled probe,the minimal detection amount ofdiseased plant total nucleic acid was approximately 400ng/spot,using DIG-labelled probe,approximately 80ng/spot;the minimal amount of CEVd-cDNA detected was about 10pg/spot with biotin hydrazide-labelled probe.Using the developed CEVd-detection kit,CEVd can be detected from diseased plants with or without symptoms.This kit is sensitive,specific,quick and easy to fol low,and has been applied in the diagnosis of viroid of diseased samples from Hunan,Hubei and Fujian provinces.

关 键 词：植物病毒 柑桔裂皮病 类病毒 生物素肼 化学标记 

分 类 号：S432.41[农业科学—植物病理学] S436.661.1[农业科学—农业昆虫与害虫防治]