地高辛的生物素-链霉亲和素免疫法测定方法的建立

Establishment of Competitive ELISA for Digoxin in Serum by Biotin avidin Technology

目的：将生物素链霉亲和素免疫反应系统引入地高辛血药浓度监测中，准确地监测地高辛血药浓度。方法：将生物素与抗ＤＩＧＩｇＧ化学连接，ＤＩＧ通过ＨＳＡ与ＨＲＰ连接，建立了生物素链霉亲和素免疫反应系统测定地高辛的竞争反应模型。结果：以０５、１、２、３、５μｇ／Ｌ五个标准点建立了竞争反应曲线，同时将ＢＮＨＳＩｇＧ作了１：４００、１：８００、１：１６００、１：３２００的稀释，探讨了竞争反应曲线的规律性变化，运用流行的四参数曲线拟合法对竞争曲线作了拟合。同时做了方法学评价。结论：流行的生物素链霉亲和素技术可以引入被动的酶联免疫吸附法中。

Objective: To develop a solid phase,competitive,enzyme immunoassay for the measurement of digoxin in serum using biotin streptavidin system.Method:Biotinyl N hydroxysuccinimide was used for introducing biotin moieties into proteins.It incorporated the biotin moiety via lysines into antibodies.The peroxidase was modified with amine enriched human serum albumin(HSA)by the addition of HSA to peroxidase,and then the periodate oxidized digoxin were at tached indirectly via the HSA to the peroxidase A model for the determination of digoxin was developed.Result:We got a standard curve for the concentrations of digoxin in serum.It was observed that standard curves had a regular change with the dilution of BNHS IgG from 1:400 to 1:3200 when the DIG HSA was diluted at 1:1200 .At the same time we fitted the curve with a four parameter function.Conclusion:Biotin Streptavidin technology can be applied to ELISA to improve the sensitivity.