柑桔黄龙病的血清学检测与诊断方法的初步研究

PRELIMARY STUDIES ON THE METHODS OF RAPID SEROLOGICAL DETECTION AND DIAGNOSIS OF THE BLO ASSOCIATED WITH CITRUS SHOOT-YELLOWING

用提纯的柑桔黄龙病病原(CYSO)免疫小鼠制备出抗体IgG,其浓度与2.7mg/ml,并证明它对CYSO具有特异性,效价测定结果:ELISA达1∶640,间接免疫荧光(IIF)达1∶1 000;斑点(?)联免疫技术(Dot-ELISA)达1∶1280;斑点免疫荧光技术(Dot-IF)达1∶1 280,检测纯化抗原(?)度,ELISA达1∶256;IIF达1∶4 096,Dot-ILISA达1∶8 192,Dot-IF达1∶32 768。ELISA只能用于检测典型的蕉柑和椪柑病材料,面IIF,dot-ILISA和Dot-IF还可用于甜橙的检测,用4种方法均不能检测出未显症材料的CYSO。

Lots of partially purified citrus yellow shoot organisms (CYSO) were used as antigen to immunize mice for the reduction of anti-CYSO ascitic fluid.The ascitic fluid proved specific to CYSO.Its titer by agar gel diffusion was 1:8 and by microprecipitation 1:512Anti-CYSO IgG was prepared by (NH4)2SO4 precipitation combined with DEAE-cellulose column chromatography.Using 1:5000 partially purified CYSO,its titer by ELISA was 1:640,by IIF was 1:1000,and that by both Dot-ELISA and Dot-IF on nitrocellulose membranes was 1:1 280.Using this IgG,a dilution of 391μg/ml of partially purified CYSO was detected by ELISA,0.24μg/ml by IIF,0.12μg/ml by Dot-ELISA and 0.031μg/ml by Dot-IF.CYSO in crude extracts of typical diseased specimens from Citrus reticulata cv.ponkon and Citrus reticalata cv tonkon could be detected comsistently by ELISA at a certain dilution,yet those from either ponkan,tankan or Citrus sinensis could be detected sensitively by either IIF,Dot-ELISA or Dot-IF.None of the symptomless specimens and those showing very early symptom could be detected as positive.