Determination of antibody titers against soluble antigen by indirect fluorescent antibody (IFA) assay using latex particles coupled with soluble antigen: A preliminary study

摘要

A method for indirect fluorescent antibody (IFA) assay for soluble antigen has been developed using latex particles (latex) as a carrier for soluble antigen. Two types of latex having grain sizes of 1.0 and 6.0 lm were used for IFA assay and were evaluated in this study. Bovine IgG and rabbit anti-bovine IgG antibody were used as soluble antigen and as primary antibody, respectively. Bovine IgG-sensitized latex on slide glass was not washed off after immersion for 45 min. IFA using latex appears to have high specificity, as no reactions between fluorescein isothiocyanate (FITC)-conjugated antibody and protein-sensitized latex were observed, and the latex did not show auto-fluorescence or non-specific reactions. Latex with a grain size of 6.0 lm was appropriate as a carrier of soluble antigen, as the amount of soluble antigen necessary for sensitization was approximately one-eighth that necessary for the small-diameter latex (1.0 lm), and large-diameter latex is easier to distinguish under the fluorescence microscope. IFA assay using soluble antigen-sensitized latex is suitable for more widespread use in the future.