免疫濁度測定及其臨床應(yīng)用課件

免疫濁度測定及其臨床應(yīng)用免疫測定IMMUNOASSAYIA免疫測定原理利用抗原抗體反應(yīng)檢測標(biāo)本中微量物質(zhì)抗原+抗體

抗原抗體復(fù)合物

Ag+AbAg*Ab免疫測定分類免疫濁度測定g~ng標(biāo)記免疫測定ng~pg免疫濁度測定免疫沉淀反應(yīng)海德堡曲線Heidelberg’sCurve免疫濁度測定分類透射濁度法散射濁度法透射濁度法和散射濁度法的光路檢測器A(透射濁度法)檢測器B(散射濁度法)I0I渾濁樣品光源散射濁度法321TimeScatterScatterSignalVersusTime

Nephelometry1.Bufferaddition2.SampleAddition3.AntibodyadditionFluidinthecellhascauseslittlescatterAddingsampleincreasesscatterslightlyAddingantibodycausesalargeincreaseinlightscattering免疫濁度測定分類2終點(diǎn)法速率法終點(diǎn)法固定時(shí)間法速率峰的測定速率峰的測定321TimeScatterMeasuretheRATEAfterAntibodyAddition

RateNephelometry1.Bufferaddition2.SampleAddition3.AntibodyadditionFirstderivative:rateofchangeofscatterovertime;

thisistheSLOPEofthecurveRateCONCENTRATIONRATE在速率峰基礎(chǔ)上完成的標(biāo)準(zhǔn)曲線抗原過剩的測定Analyte-antibodycomplexingAnalyteAntibody

AntigenExcessRATEofSCATTER抗體過量時(shí)的反應(yīng)抗原過量時(shí)的反應(yīng)60850反應(yīng)時(shí)間(秒)ARRAY抗原過量自動檢測加入校正液藥物濃度測定免疫濁度抑制測定(小分子物質(zhì))抗體試劑的要求1特異性2親和力3反應(yīng)時(shí)保持抗體過剩測定范圍應(yīng)包括高于及低于正常值的50%5測定的精密度應(yīng)在5%左右校正曲線：聚乙二醇的增強(qiáng)作用抗原濃度C1C2C3吸光度4%聚乙二醇不加聚乙二醇試劑組成配方舉例試劑1100mmolTris或磷酸緩沖液pH7.2~8.24%聚乙二醇(分子量6000或10000)1%非離子型表面活性劑150mmol氯化鈉或硫酸鈉0.1g/l疊氮鈉試劑2100mmolTris或磷酸緩沖液pH7.2~8.220%多克隆羊抗血清150mmol氯化鈉0.1g/l疊氮鈉抗原濃度測定范圍抗體過?？乖^剩Ag*Ab技術(shù)特性并行的檢測系統(tǒng):RateNIPIARateNephelometryNIPIA：近紅外顆粒速率法免疫分析BenefitsAbilitytoexpandthemenuintohighsensitivityassaysincluding:Ferritin,Digoxin,andIgE.NIReliminatestheinterferenceofbiologicalsubstancessuchasbiliribinandhemoglobin.NIPIA：近紅外顆粒速率法免疫分析優(yōu)點(diǎn)擴(kuò)展檢測菜單，增加高敏項(xiàng)目分析：如鐵蛋白，地高辛，IgE，超敏CRP等近紅外波長分析消除生物物質(zhì)如膽紅素和血色素的干擾NIR：940nm波長近0oC檢測角檢測顆粒大小：200~500nm直徑速率法測定NIPIA：940nm