摘要
以改进的直接组织斑免疫测定(IDDTB)的直接法,用碱性磷酸酯酶(Ap)或辣根过氧化物酶(HRP)标记的单克隆抗体(IgG)检测病组织中的马铃薯X病毒(PVX)、菁花叶病毒(TuMV)和马铃薯青枯细菌均获得了满意的结果。该法明显地比酶联免疫吸附测定(ELISA)和圆点免疫结合测定(DIBA)的试验程序简单、快速和准确可靠,整个检测程序可在1.5～2小时内完成。据推算其检测PVX的灵敏度相当于0.625ng/ml,可检测病叶的最低稀释度相当于1:320。以IDDTB的间接法用Ap标记的羊抗兔IgG或HRP标记的鼠抗免IgG以及兔抗病毒和细菌的多克隆抗体(IgG)检测病组织中的烟草花叶病毒(TMV)、黄瓜花叶病毒(CMV)、马铃薯卷叶病毒(PLRV)和马铃薯环腐细菌,除耗时比上述直接法稍长外(约3小时),同样获得较满意的结果。两法均适于在田间应用。IDDTB直接法和间接法检测结果的可靠性均由免疫电镜法得到证实。上述免疫反应的结果因底物不同呈现不褪色的蓝色或深紫色,极易同健康叶片呈现的绿色相区别,并可在室温条件下长期保存。
The results of improved direct Immunological Detection of Direct Tissue Blotting(IDDTB)
detecting potato virus X (PVX ),turnip mosaic virus(TuMV )and bacteria wilt(BW)(pseu-
domonas solanacearum E.F.Smith)in infected tissues with Alkaline phosphatase(Ap)or
Horseradish peroxidase(HRP)conjugated monoclonal antibody(IgG)were satisfactory.The pro5cedure was simple,rapid and reliable compared with ELISA and DIBA,and could be finished in 61.
～2 hours.The sensitivities were calculated by 0.
25ng/ml for detecting purified PVX and
1:320 for detecting infected leaves.The results of improved indirect IDDTB detecting tobacco
mosaic virus(TMV),cucumber mosaic virus(CMV),potato leafroll virus(PLRV)and ring —rot
(RR)(Erwinia carotorora Subspp)in infected tissues with Ap conjugated goat anti rabbit IgG or
HRP conjugated mouse anti rabbit IgG and rabbit anti viruses or polyclonal antibodies to bacteria
were also satisfactory except that detecting time was longer than the IDDTB(about 3 hours).
The reliability of detective results of two methods were certified by Immunosorbent Electron Mi
croscope(ISEM).
Unfade blue and deep purple produced by the reaction of enzymes and substrates were easily
differentiated from the green of health leaves,and could be preserved for a long time in room
temperature.
The results of improved direct Immunological Detection of Direct Tissue Blotting(IDDTB)
detecting potato virus X (PVX ),turnip mosaic virus(TuMV )and bacteria wilt(BW)(pseu-
domonas solanacearum E.F.Smith)in infected tissues with Alkaline phosphatase(Ap)or
Horseradish peroxidase(HRP)conjugated monoclonal antibody(IgG)were satisfactory.The pro5cedure was simple,rapid and reliable compared with ELISA and DIBA,and could be finished in 61.
～2 hours.The sensitivities were calculated by 0.
25ng/ml for detecting purified PVX and
1:320 for dete
出处
《植物病理学报》
CAS
CSCD
北大核心
1993年第4期367-371,共5页
Acta Phytopathologica Sinica
