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혈중 변역 복합체 검출을 위한 Raji Cell ELISA법 의 개발
임태숙 ( T S Rim ),이은미 ( E M Lee ) 대한임상검사과학회 1991 대한임상검사과학회지(KJCLS) Vol.23 No.1
There are various kinds of circulating immune complexes(cic) and we have a number of methods of measuring them. However, any of them can not alone measure cic correctly, usually two or three methods are combined to be applied. Currently, in the immunological laboratory, the solid-phase Clq ELISA (Enzyme Linked Immuno -sorbent Assay) which uses a component of the complement system(C1q) is commonly used, but in an effort to more correctly measure cic, another kind of ELISA using Raji cell was experimented. To be more specific, optimum conditions were put forward synthesizing the reaction factors of the Raji cell ELISA and based on those conditions, cics of the autoimmune disease patients and the normal control group were measured with the three methods (Raji cell ELISA, Raji cell RIA and Clq ELISA). Following results were obtained. 1. The optimum conditions of the Raji cell ELISA which displayed the best standard curve were ; 1 : 4 dilution ratio of AHGG. 1 : 4 dilution ratio of NHS, 2 X l06cells/ml of Raji cell numbersㆍ 1 : 1, 000 dilution ratio of conjugate, 20minutes of substrate (PNP) reaction time and 4ㆍc reaction temperature of sample and Raji cells. 2. The coefficient (C. V.) variation of interassays of the Raji cell ELISA was 13.19%, which that of intraassay was 5.16%. 3. For the normal control group the mean of cic levels was 10.3±5.0(mean±S.D),ug of AHGG equivalent/ml with the Raji cell RIA and 11. 7±5. O,ug of AHGG equivalent/ml respectively. 4. For the autoimmune disease patient group, the mean of cic levels was 89. 3 ± 105 .l(mean ± S. D.) ,ug of AHGG equivalent/ml with. the solid -phase C1q ELISA, 118.5±140.0,ug of AHGG equivalent/ml with the Raji cell RIA and 195. 5± 257. 5,ug of AHGG equivalent/ml with the Raji cell ELISA, all of which showed higher levels of cics than the normal control group significantly. 5. It was also revealed that correlation between the solid-phase Clq ELISA and Raji cell ELISA was 0.34(P<0.0001), while the correlation between the solid-phase C q ELISA and the Raji cell RIA and between the Raji cell RIA and the Raji cell ELISA were 0. 49 (P < 0. 0001) and 0. 63 (P < 0. 0001) respectively. Summing up, the optimum condition of Raji cell ELISA is established, and in order to more correctly measure the cic levels, the principle of measurement and the sensitivity of the tests should be considered. In this regard, the combined use of the solid-phase Clq ELISA and the newly-experi Raji cell ELISA is believed to be more effective than single C1 q ELISA.