Abstract
In the present study, quantitative immunoelectrophoresis was conducted for TMV estimation, by using antibody conjugated with fluorescein isothiocyanate (FITC). FITC-conjugated antibody (FA) showed the same properties as non-conjugated antibody, in precipitate formation to antigen, stainability with Amido Black 10-B and electrophoretic mobility of antigen-antibody complex. Using broad ranges of TMV and FA concentrations, optimal conditions for TMV estimation were statistically clarified in each combination, and the linear regression line was offered within defined range of TMV to each FA concentration for a basis of routine quantification of TMV. Sensitivity for detection was highly enhanced by using FA. TMV as small as 0.008μg was successfully detected only by fluorescence of conjugating FITC emitted under UV-irradiation. The high accuracy of TMV quantification was reached when applied to inoculated or systemically infected tomato leaves. In one case, the variability of the migration distance was below 3% level and that of calculated TMV contents was between 5% and 8% level.
