효소면역측정법을 위한 장티푸스 균체항원의 부착방법 = Methods for Coating the Killed Whole Cell Antigens of Salmonella typhi in Enzyme - linked Immunosorbent Assay

다국어 초록 (Multilingual Abstract)

The advantages of enzyme-linked immunosorbent assay(ELISA) are its senstivity and simplicity in detecting IgM and IgA antibody. To apply ELISA to diagnosis of typhoid fever, antigen such as lipopolysacc-haride of Salmonella typlii or killed whole cell must be coated on solid phase. It is easy to coat lipopolysaccharide on ELISA plate but troublesome to purify it. As it is easy to obtain the killed whole cells, the development of the appropriate method by which those antigens of S. typhi are optimally coated on solid phase is needed.
To establish the appropriate method, carbonate buffer, methanol or poly-L-lysine was applied as binding substance on polystyrene or polyvinylchloride plate as solid phase when the killed whole cell antigens of .S. typhi varided as follows: 10_6, 10_7, 10_8 and 10_9 cell/ml.
The criteria of the optimal method were determined as follows: 1. The optical density of positive sera is above 1.0(0.6 in IgM) at 1: 10 serum dilution and is 0.3(0.2 in IgM) higher than that of negative sera: 2. The C3.D. of sera is flat or lowering according to serum dilution; 3. It must be that the O.D. of negative sera is lower than 0.2 at the point of serum dilution where the O.D. of positive sera is higher than 1.0 (0.5 in IgM).
The results obtained were summarized as follows;
1. The methods which fitted the above criteria were to use poly-L-lysine as binding substance, polyvinylchloride plate as solid phase and 10 cell/ml as antigen concentration of Z typhi (poly-L- lysine/ polyvinylchloride/10 and poly-L-lysine/polyvinylchloride/10 in detecting IgG antibody, methanol/ po-lystyrene/10_9, poly-L-lysine /polyvinylchloride/10_8 and poly-L-lysine /pilyvinylchloride /10_9 in IgM and carbonate buffer/polystyrene/10, carbonate buffer/polystyrene/10_8, methanol/polystyrene/10_5, methanol/ polyvinylchloride/10_9, methanol/polyvinylchloride/10_9, poly-L-lysine/polyvinylchloride/10_9 and poly-L-1ysine/polyvinylchloride/10_9 in IgA.
2. The coating method using poly-L -lysine, polyvinylchloride plate and 10_9 cell/ml was best toassay gG, IgM and IgA antihcody all in one. By this method, to assay the each immunoglobulin calss with an appropriate fixed serum dilution, 1: 320 dilution was best.