免疫浊度测定及其临床应用

Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,Edited by E. Dalla Dea ESTS,免疫浊度测定,及其临床应用,免 疫 测 定,IMMUNOASSAY,I A,免疫测定原理,利用抗原抗体反应检测标本中微量物质,抗原+抗体,抗原抗体复合物,Ag +Ab Ag * Ab,免疫测定特点,特异性,抗原性不同的物质不干扰,敏感性,g-ng-pg,可测物,蛋白质，酶，激素，药物 ，,毒品,免疫测定分类,免疫浊度测定,g ng,标记免疫测定,ng pg,免疫浊度测定,免疫沉淀反应,海德堡曲线,Heidelbergs Curve,抗原浓度,测定范围,抗体过剩,抗原过剩,Ag * Ab,免疫浊度测定分类,透射浊度法,散射浊度法,透射浊度法和散射浊度法的光路,检测器,A,(透射浊度法),检测器,B,(散射浊度法),I,0,I,浑浊,样品,光源,散射浊度法,Analyte Antibody Other stuff,Analyte-antibody complexing,Now big enough to scatter light,Too small to detect optically,What happens inside the flow cell?,3,2,1,Time,Scatter,Scatter Signal Versus Time,Nephelometry,1.,Buffer addition 2. Sample Addition 3. Antibody addition,Fluid in the cell has causes little scatter,Adding sample increases scatter slightly,Adding antibody causes a large increase in light scattering,免疫浊度测定分类2,终 点 法,速 率 法,终 点 法,固定时间法,速率散射浊度法,速率峰的测定,速率峰的测定,3,2,1,Time,Scatter,Measure the RATE After Antibody Addition,Rate Nephelometry,1.,Buffer addition 2. Sample Addition 3. Antibody addition,First derivative: rate of change of scatter over time;this is the SLOPE of the curve,Rate,PEAK RATE,TIME,RATE,Rate of Change of Scatter Signal versus Time,CONCENTRATION,RATE,在速率峰基础上完成的标准曲线,抗原过剩的测定,Analyte-antibody complexing,Analyte Antibody,Antigen Excess,RATE of SCATTER,抗体过量时的反应,抗原过量时的反应,60,85,0,反应时间(秒),ARRAY,抗原过量自动检测,加入校正液,药物浓度测定,免疫浊度抑制测定,(小分子物质),1,2,Conjugate-antibody complexing,2,1,3,Inhibition of complexing by hapten (drug),1.,Conjugate (hapten on carrier)2. Antibody3. Hapten (drug),免疫浊度测定试剂,抗体试剂的要求,1 特异性,2 亲和力,3 反应时保持抗体过剩,测定范围应包括高于及低于,正常值的50%,5 测定的精密度应在5%左右,校正曲线：聚乙二醇的增强作用,抗原浓度,C1,C2,C3,吸光度,4%聚乙二醇,不加聚乙二醇,试剂组成配方举例,试剂1,100,mmol Tris,或磷酸缓冲液,pH7.2 8.2,4%,聚乙二醇(分子量6000或10000),1%非离子型表面活性剂,150,mmol,氯化钠或硫酸钠,0.1,g/l,叠氮钠,试剂2,100,mmol Tris,或磷酸缓冲液,pH7.2 8.2,20%,多克隆羊抗血清,150,mmol,氯化钠,0.1,g/l,叠氮钠,抗原浓度,测定范围,抗体过剩,抗原过剩,Ag * Ab,技术特性,并行的检测系统:,Rate NIPIA,Rate,Nephelometry,NIPIA：,近红外颗粒速率法免疫分析,Benefits,Ability to expand the menu,into high sensitivity assays,including: Ferritin, Digoxin, and IgE.,NIR eliminates the interference,of biological substances,such as biliribin and hemoglobin.,NIPIA：,近红外颗粒速率法免疫分析,优点,扩展检测菜单，增加高敏项目分析：,如铁蛋白，地高辛,，,IgE，,超敏,CRP,等,近红外波长分析消除生物物质如胆,红素和血色素的干扰,NIR：940nm,波长,近0,o,C,检测角,检测颗粒大小：200500,nm,直径,速率法测定,NIPIA,： 940nm,LED,光源,速率散射法,：670,nm,激光光源,蛋白应用分类,心血管功能监测,载脂蛋白,A1,载脂蛋白,B,脂蛋白,a,