罗汉果花叶病毒(WMV-2-Luo)抗血清制备及ELISA检测技术研究  

Polyclonal antiserum preparation and ELISA detection of Luohanguo mosaic virus(WMV-2-Luo)

作　　者：秦碧霞[1] 蔡健和[1] 刘志明[1] 余玉冰[1] 

机构地区：[1]广西农业科学院植物保护研究所,广西南宁530007

出　　处：《西南农业学报》2001年第4期14-16,共3页Southwest China Journal of Agricultural Sciences

基　　金：广西青年科学基金资助项目 (桂科青 972 10 12 )

摘　　要：用西葫芦作为罗汉果花叶病毒 (WMV 2 Luo)的繁殖寄主 ,通过差速离心和PEG二次沉淀法得到WMV 2 Luo的提纯液 ,用该病毒提纯液免疫日本大白兔 ,获得了WMV 2 Luo兔抗血清。经ELISA测定 ,其效价为 1/5 12 0 ,以 0 1mol/L磷酸盐缓冲液 (pH7 8,含 1 5 %NaDIECA)作为样品制备缓冲液 ,用间接ELISA法检测WMV 2 Luo的灵敏度为 1/32 0 0。用此方法检测罗汉果组培苗带毒情况和田间病株均获得成功。Luohanguo mosaic virus (WMV-2-Luo) were purified from Cucurbit pepo infected by WMV-2-Luo,and concentrated by one cycle of differential centrifugation and PEG double precipitation.An antiserum against WMV-2-Luo with a virus-specific titre of 1/5120 by ELISA was obtained by immunizing a Janpanese rabbit using the purified virus preparation.With a proper phosphate buffer solution,i.e.0.1 mol/L phosphate buffer(pH7.8,including 1.5%Sodium diethyldityiocarbamate)for preparing the crude plant sap to be detected,it was possible to increase the sensitivity of DAC-ELISA to a dilution of plant sap at a rate of 1/3200.It is successfully to detect WMV-2-Luo of the samples from tissue cultural planted and field.

关 键 词：罗汉果花叶病毒 提纯 抗血清制备 间接ELISA法 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]