全自动化学发光免疫分析仪运行原理课件.ppt

1、全自动化学发光免疫分析仪运行原理化学发光基本原理化学发光基本原理v 反应体系中的某种物质的分子反应体系中的某种物质的分子(反应物、反应物、产物、中间体或荧光物质产物、中间体或荧光物质)吸收了反应所释吸收了反应所释放的能量而由基态跃迁至激发态，然后再放的能量而由基态跃迁至激发态，然后再从激发态返回基态，同时将能量以光辐射从激发态返回基态，同时将能量以光辐射的形式释放出来，产生化学发光的形式释放出来，产生化学发光.v将化学发光反应应用于分析化学，根据某将化学发光反应应用于分析化学，根据某一时刻的发光强度或反应的发光总量来确一时刻的发光强度或反应的发光总量来确定体系的相应组分含量的分析方法叫化学定体

2、系的相应组分含量的分析方法叫化学发光分析法发光分析法.化学发光的反应必须具备两个条件化学发光的反应必须具备两个条件v一、是该反应能释放出一定的能量，且释放出的一、是该反应能释放出一定的能量，且释放出的能量可以被某种反应产物或中间体所吸收，使之能量可以被某种反应产物或中间体所吸收，使之处于激发态；处于激发态；v二、是这种激发态产物应具有一定的化学发光量二、是这种激发态产物应具有一定的化学发光量子产率，或者可以将其能量有效地转移给某种荧子产率，或者可以将其能量有效地转移给某种荧光物质，产生光辐射光物质，产生光辐射.v基于此，并不是任何化学反应都能产生化学发光基于此，并不是任何化学反应都能产生化学发

3、光反应，因此，如果测量条件适当，化学发光分析反应，因此，如果测量条件适当，化学发光分析会有足够的选择性会有足够的选择性.化学发光的分类化学发光的分类v化学发光反应参与的免疫测定分为二种类型。化学发光反应参与的免疫测定分为二种类型。v第一种是以发光剂作为酶免疫测定的底物，通过发第一种是以发光剂作为酶免疫测定的底物，通过发光反应增强测定的敏感性。光反应增强测定的敏感性。如：化学发光酶免疫测定（如：化学发光酶免疫测定（CLEIA，与酶标，与酶标ELISA法类法类似，即最后一步酶反应所用底物为发光剂似，即最后一步酶反应所用底物为发光剂）v第二种是以发光剂作为抗体或抗原的标记物，直接第二种是以发光剂作为

4、抗体或抗原的标记物，直接通过发光反应检测标本中抗原或抗体的含量。通过发光反应检测标本中抗原或抗体的含量。如：化学发光标记免疫测定亦称化学发光免疫测定如：化学发光标记免疫测定亦称化学发光免疫测定（CLIA）、电化学发光免疫测定（）、电化学发光免疫测定（ECLI）ABBOTT i 2000SR 结构结构v1.i 2000SR processing modulev2.RSH(retest sample handler)v3.SCC(system control center)ABBOTT i 2000SR 结构结构v1.Front processing center coverv2.Processi

5、ng module keypadv3.Supply and waste center doorv4.Card cage doorv1.Rear processing center coverv2.Rear processing center access panelv3.Power supply panelv4.Pump bay panelABBOTT i 2000SR 结构结构1.Sample hardware components:Provide sample aspiration and dispense.2.2.Reagent hardware components:Provide r

6、eagent aspiration and dispense.3.3.Process path hardware components:Position the RVs for sample and reagent aspiration,mixing,washing,and CMIA processingv1.Sample pipettorv STAT pipettor1.Sample and STAT pipettors(S and ST):2.2.Sample and STAT syringes(SS and STS):3.3.Sample and STAT wash stations(S

7、W and STW):1.Reagent carousel2.2.Reagent bar code reader3.3.Reagent pipettors4.4.Reagent syringes 5.5.Reagent wash stations1.Load diverter 2.RV access door 3.RV loader and hopper assembly 4.STAT diverter5.Vortexers(Mix)6.Wash zone diverter 7.Wash zone manifolds(WZ1,WZ2):8.Process path drive motor 9.

8、Pre-trigger/trigger manifold 10.CMIA reader(CMIA)11.Liquid waste arm 12.RV unloaderABBOTT i 2000SR检测原理检测原理vArchitect I系统采用化学发光微粒子免疫分析技术系统采用化学发光微粒子免疫分析技术（Chemiluminesent Microparticle ImmunoAssay,CMIA）检测样品中的抗原，抗体和分析物。检测样品中的抗原，抗体和分析物。v即：磁性微粒子包被的捕捉分子（抗原，抗体或病毒颗粒）即：磁性微粒子包被的捕捉分子（抗原，抗体或病毒颗粒）特异的与被分析物中抗原、抗体结

9、合形成抗原抗体复合物，特异的与被分析物中抗原、抗体结合形成抗原抗体复合物，再与吖啶酯标记的连接物反应形成双抗体夹心抗原抗体复合再与吖啶酯标记的连接物反应形成双抗体夹心抗原抗体复合物。吖啶酯在过氧化氢的稀碱溶液中发生氧化还原反应生成物。吖啶酯在过氧化氢的稀碱溶液中发生氧化还原反应生成10-甲基吖啶酮，当它恢复到基态时发光，根据发光强度可甲基吖啶酮，当它恢复到基态时发光，根据发光强度可计算出分析物的浓度计算出分析物的浓度。ABBOTT i 2000SR试剂组成试剂组成v试剂组成试剂组成 v中圈（中圈（RI）：包被有：包被有抗体的顺磁性微粒抗体的顺磁性微粒子子v内圈（内圈（R2）：吖啶：吖啶酯包被的

10、抗体酯包被的抗体v外圈外圈：样本稀释液：样本稀释液v预激发液预激发液：1.32%的的过氧化氢过氧化氢v激发液激发液：0.35mol./L的氢氧化钠的氢氧化钠v清洗缓冲液清洗缓冲液：磷酸：磷酸盐缓冲液盐缓冲液R1试剂试剂R2试剂试剂ABBOTT i 2000SR反应类型反应类型v1.双抗原（抗体）夹心一步法双抗原（抗体）夹心一步法v2.双抗原（抗体）夹心两步法双抗原（抗体）夹心两步法 Assay processing for One step 25 Assay processing for Two step 18-4 STAT assay processing for One step 11 S

11、TAT assay processing for Two step 4-4Assay processing for Two step 18-4(i System)v1.At position 1 the sample pipettor dispenses the sample into the RV(reaction vessel).v2.At position 2 the R1 pipettor dispenses the microparticles.v3.At position 3 the vortexer mixes the sample and microparticles.v4.A

12、t positions 4-63 the reaction mixture incubates for 18 minutes.共共112个个RV杯空位，每杯空位，每18秒秒转一个空位，转一个空位，200T/h。常规。常规28Min/TESTAssay processing for Two step 18-4(i System)v5.At positions 64-67 the wash zone 1 manifold washes the reaction mixture in the RV,and then removes unbound materials Assay processing

13、 for Two step 18-4(i System)v6.At position 71 the R2 pipettor dispenses acridinium-labeled conjugate.v7.At position 72 the vortexer mixes the reaction mixture.v8.At positions 73-86 the reaction mixture incubates for 4 minutes.Assay processing for Two step 18-4(i System)v9.At positions 87-90 the wash

14、 zone 2 manifold washes the reaction mixture in the RV,and then removes unbound materials.Assay processing for Two step 18-4(i System)v10.At position 94 the pre-trigger nozzle dispenses Pre-Trigger Solution to the reaction mixture,and then mixes using the vortexer.v11.At position 98 the CMIA optical

15、 system takes a background read,the trigger nozzle dispenses Trigger Solution to the reaction mixture,and then the CMIA optical system takes an activated read.v12.At position 100 the liquid waste arm aspirates the liquid waste from the RV.v13.At position 109 the RV unloader removes the RVAssay proce

16、ssing for One step 25(i System)v1.At position 1 the sample pipettor dispenses the sample into the RV(reaction vessel).v2.At position 2 the R1 pipettor dispenses the microparticles and acridinium-labeled conjugate.vNOTE:For a delayed one-step assay the R2 pipettor adds the acridinium-labeled conjugat

17、e at position 71 and the vortexer mixes the reaction mixture at position 72.v3.At position 3 the vortexer mixes the sample,microparticles,and conjugate.v4.At positions 4-86 the reaction mixture incubates for 25 minutes.Assay processing for One step 25(i System)v5.At positions 87-90 the wash zone 2 m

18、anifold washes the reaction mixture in the RV,and then removes unbound materials.vNext position is same to the two step18-4.STAT assay processing for One step 11(i System)v1.At position 47 the STAT pipettor dispenses the sample into the RV(reaction vessel).v2.At position 48 the R2 pipettor dispenses

19、 the microparticles and acridinium-labeled conjugate.v3.At position 49 the vortexer mixes the sample,microparticles,and conjugate.v4.At positions 50-86 the reaction mixture incubates for 11 minutes.STAT assay processing for One step 11(i System)v5.At positions 87-90 the wash zone 2 manifold washes t

20、he reaction mixture in the RV,and then removes unbound materials.vNext position is same to the two step18-4.STAT assay processing for Two step 4-4(i System)v1.At position 47 the STAT pipettor dispenses the sample into the RV(reaction vessel).v2.At position 48 the R2 pipettor dispenses the microparti

21、cles.v3.At position 49 the vortexer mixes the sample and microparticles.v4.At positions 50-63 the reaction mixture incubates for 4 minutes.STAT assay processing for Two step 4-4(i System)v5.At positions 64-67 the wash zone 1 manifold washes the reaction mixture in the RV,and then removes unbound mat

22、erials.STAT assay processing for Two step 4-4(i System)v6.At position 71 the R2 pipettor dispenses acridinium-labeled conjugate.v7.At position 72 the vortexer mixes the reaction mixture.v8.At positions 73-86 the reaction mixture incubates for 4 minutes.STAT assay processing for Two step 4-4(i System)v9.At positions 87-90 the wash zone 2 manifold washes the reaction mixture in the RV,and then removes unbound materials.vNext position is same to the two step18-4.ABBOTTi2000SR操作注意事项v1、库存（BUF、RV杯）保证充足。v2、标本预处理（纤维、气泡、溶血、脂浊、浑浊、条码、摆放位置、项目是否输入）。v3、报警知识的学习及处理（代码、关键词、描述及处理方法）。谢谢！谢谢！