应用DIBA检测植物病毒的研究

 点免疫结合测试法(Dot Immunobinding Assay,DIBA)检测提纯TMV、PVY、CMV的最低检出率分别达到0.184ng/ml、1.28×10³ng/ml、0.056ng/ml。检测感病叶片粗汁液的最高稀释度依次为1:819,200、1:102,400、1:819,200。其中,检测TMV的最低检出率及最高稀释度分别比Hibi(1985)1]的报道高544倍和100余倍。检测PVY粗汁液的最高稀释度比Banttari(1985)的报道高15倍2]。HRP-IgG、抗血清在所使用的稀释度范围内(1:1000-8000、1:500-4000)对结果没有明显影响。但检测PVY时,随着血清稀释度的提高最低检出率明显下降。利用HRP-A代替HRP-IgG可以获得更为理想的检测效果。改进的DIBA(暂称为DSA-DIBA)检测TMV时未发现有任何假阳性反应。DIBA检测PVY、TuMV、PRSV-T揭示了它们相互间有效远缘的血清学相关性。实验表明,DIBA是检测植物病毒专化、快速、简便、灵敏、经济的新方法。

ON THE DETECTION OF PLANT VIRUSES BY DIBA

The recently developed method of dot-immunobinding assay(DIBA) was adopted in this experiment to detect three plant viruses of different mor-phology.The purified TMV was satisfactorily detected by DIBA at 0.184ng/ml,purified PVY at 1.28x10³ ng/ml and purified CMV at 0.056ng/ml,requiring about 5 hours to finish a test.Also the sap of tobacco plants infected with TMV,PVY and CMV respectively were detected by DIBA at dilutions of 1:819,200,1:102,400 and 1:819,200 respectively.Since the background and nonspecific reactions are critical problems in DIBA,the use of HRP-A resulted in no background color and less nonspecific reac-tions than in the case of using HRP-IgG.Therefore it is of the opinion that to select appropriate dilutions of antiserum and healthy leaves are also important.The present experiments showed that DIBA inherited almost all advantages of ELISA,yet more sensitive,economic,rapid and simpler than the latter.The writers prefer to use DIBA rather than ELISA to detect and identify plant viruses.