应用免疫电镜吸附双修饰法检测植物病毒

COMBINED IMMUNOSORBENT AND DOUBLE DECORATION METHOD FOR DETECTING PLANT VIRUSES BY ELECTRON MICROSCOPY

免疫电镜技术是目前最为灵敏的血清学技术之一。该技术所需抗原和抗体均很微量,操作时间短,准确性高,可以直接观察与抗体结合的病毒,深受植物病毒研究工作者的欢迎。1981年,Kerlan等建立了一种称作免疫电镜双修饰法的新技术。该技术由于在处理过程中增用了羊抗兔免疫球蛋白而加强了修饰效果,双修饰后的病毒粒体在放大700～2500倍下即可看见。本文报道利用该技术成功地检测了烟草花叶病毒(TMV)、黄瓜花叶病毒(CMV)、马铃薯Y病毒(PVY),获得了很高的灵敏度和明显的修饰效果。

By the method of combined immunosorbent and double decoration, the purified TMV was able to be detected at 0.184 ng/ml, PVY at 0.128ng/ml and CMY at 0.056ng/ml by electron microscopy. The crude saps of tobacco plants infected with TMV, PVY and CMV respectively which were diluted to 1 : 819,200, 1 : 1,600 and l : 200,000 were also readily detected by this method. It needs only one hour and several microliters of antigen and antiserum to detect the doubly decorated viruses with electron microscopy at an magnification as low as 2,000 times and it is therefore also possible to be observed with high power optical microscope.