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효소면역측정법을 위한 장티푸스 균체항원의 부착방법
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저자명
김윤원,황응수,국윤호,최강원,김익상,차창용,이승훈,Kim. Youn-Won,Hwang. Eung-Soo,Kook. Yoon-Hoh,Choi. Kang-Won,Kim. Ik-Sang,Cha. Chang-Yong,Lee. Seung-
간행물명
대한미생물학회지
권/호정보
1985년|20권 1호|pp.91-102 (12 pages)
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대한미생물학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

The advantages of enzyme-linked immunosorbent assay(ELISA) are its senstivity and simplicity in detecting IgG, IgM and IgA antibody. To apply ELISA to diagnosis of typhoid fever, antigen such as lipopolysaccharide of Salmonella typhi 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 O.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 $10^7;cell/ml$ as antigen concentration of S. typhi(poly-L-lysine/polyvinylchloride/$10^7$) and poly-L-lysine/polyvinylchloride/$10^8$ in detecting IgG antibody, methanol/polystyrene/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgM and carbonate buffer/polystyrene/$10^8$, carbonate buffer/polystyrene/$10^9$, methanol/polystyrene/$10^8$, methanol/polyvinylchloride/$10^8$, methanol/polyvinylchloride/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgA. 2. The coaling method using poly-L-lysine, polyvinylchloride plate and $10^8;cell/ml$ was best to assay IgG, IgM and IgA antibody all in one. By this method, to assay the each immunoglobulin calss with an appropriate fixed serum dilution, 1:320 dilution was best.