全自動化學發(fā)光免疫分析儀運行原理-PPT精選課件

1、美國雅培i2000化學發(fā)光免疫分析儀吉林市中心醫(yī)院滕彥玲化學發(fā)光基本原理 反應體系中的某種物質的分子(反應物、產(chǎn)物、中間體或熒光物質) 吸收了反應所釋放的能量而由基態(tài)躍遷至激發(fā)態(tài)，然后再從激發(fā)態(tài)返回基態(tài)，同時將能量以光輻射的形式釋放出來，產(chǎn)生化學發(fā)光.將化學發(fā)光反應應用于分析化學，根據(jù)某一時刻的發(fā)光強度或反應的發(fā)光總量來確定體系的相應組分含量的分析方法叫化學發(fā)光分析法. 化學發(fā)光的反應必須具備兩個條件一、是該反應能釋放出一定的能量，且釋放出的能量可以被某種反應產(chǎn)物或中間體所吸收，使之處于激發(fā)態(tài)；二、是這種激發(fā)態(tài)產(chǎn)物應具有一定的化學發(fā)光量子產(chǎn)率，或者可以將其能量有效地轉移給某種熒光物質，產(chǎn)生光輻

2、射. 基于此，并不是任何化學反應都能產(chǎn)生化學發(fā)光反應，因此，如果測量條件適當，化學發(fā)光分析會有足夠的選擇性. 化學發(fā)光現(xiàn)象的發(fā)現(xiàn)最早發(fā)現(xiàn)的化學發(fā)光現(xiàn)象發(fā)生在生物體內，即熒火蟲，現(xiàn)在稱之為生物發(fā)光(Bioluminescence). 到了十九世紀后期人們發(fā)現(xiàn)簡單的非生物有機化合物也能產(chǎn)生化學發(fā)光.1877年，發(fā)現(xiàn)洛汾堿(2，4，5-三苯基咪唑)在堿性介質中被過氧化氫等試劑氧化時發(fā)出綠色的光. 1928年，觀察到魯米諾(3-氨基苯二甲酰肼)在堿性介質中的 化學發(fā)光行為. 1935年，第一個報告了光澤精(N,N-二甲基二吖啶硝酸鹽)與過氧化氫反應產(chǎn)生化學發(fā)光.到現(xiàn)在的吖啶酯、三聯(lián)吡啶釕等發(fā)光標記物應

3、用技術的成熟。化學發(fā)光分析法的特點由于化學發(fā)光分析不使用任何光源，避免了背景光和雜散光的干擾，降低了噪聲，大大提高了信噪比，因而，化學發(fā)光分析法一般都有很高的靈敏度，通?？蓽y定納克級或皮克級的化學成分.測量化學發(fā)光強度，多采用光電轉換裝置，用函數(shù)記錄儀或數(shù)顯裝置，記錄化學發(fā)光的相對強度. 以最大發(fā)光強度(曲線的峰高)定量被測組分的濃度. 由于流動注射分析(FIA)技術的發(fā)展，流動注射化學發(fā)光儀也廣泛使用. 配備微機系統(tǒng)，自動進樣，儲存記錄，打印結果，使化學發(fā)光分析的速度更快.化學發(fā)光的分類化學發(fā)光反應參與的免疫測定分為二種類型。第一種是以發(fā)光劑作為酶免疫測定的底物，通過發(fā)光反應增強測定的敏感性

4、。如：化學發(fā)光酶免疫測定（CLEIA，與酶標ELISA法類似，即最后一步酶反應所用底物為發(fā)光劑 ） 第二種是以發(fā)光劑作為抗體或抗原的標記物，直接通過發(fā)光反應檢測標本中抗原或抗體的含量。 如：化學發(fā)光標記免疫測定亦稱化學發(fā)光免疫測定（CLIA）、電化學發(fā)光免疫測定（ECLI） ABBOTT i2000SR 結構1. i2000SR processing module2. RSH (retest sample handler)3. SCC (system control center) ABBOTT i2000SR 結構1. Front processing center cover2. Proc

5、essing module keypad3. Supply and waste center door4. Card cage door1. Rear processing center cover2. Rear processing center access panel3. Power supply panel4. Pump bay panelABBOTT i2000SR 結構Sample hardware components: Provide sample aspiration and dispense.2. Reagent hardware components: Provide r

6、eagent aspiration and dispense.3. Process path hardware components: Position the RVs for sample and reagent aspiration, mixing, washing, and CMIA processing1. Sample pipettor STAT pipettorSample and STAT pipettors (S and ST): 2. Sample and STAT syringes (SS and STS): 3. Sample and STAT wash stations

7、 (SW and STW):Reagent carousel2. Reagent bar code reader3. Reagent pipettors4. Reagent syringes 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

8、motor 9. Pre-trigger/trigger manifold 10. CMIA reader (CMIA)11. Liquid waste arm 12. RV unloaderABBOTT i2000SR檢測原理Architect I系統(tǒng)采用化學發(fā)光微粒子免疫分析技術（Chemiluminesent Microparticle ImmunoAssay, CMIA）檢測樣品中的抗原，抗體和分析物。即：磁性微粒子包被的捕捉分子（抗原，抗體或病毒顆粒）特異的與被分析物中抗原、抗體結合形成抗原抗體復合物，再與吖啶酯標記的連接物反應形成雙抗體夾心抗原抗體復合物。吖啶酯在過氧化氫的稀堿溶液

9、中發(fā)生氧化還原反應生成10-甲基吖啶酮，當它恢復到基態(tài)時發(fā)光，根據(jù)發(fā)光強度可計算出分析物的濃度 。ABBOTT i2000SR試劑組成試劑組成 中圈（RI）：包被有抗體的順磁性微粒子內圈（R2）：吖啶酯包被的抗體外圈：樣本稀釋液預激發(fā)液：1.32%的過氧化氫激發(fā)液：0.35mol./L的氫氧化鈉清洗緩沖液：磷酸鹽緩沖液R1試劑R2試劑ABBOTT i2000SR反應類型1.雙抗原（抗體）夾心一步法2.雙抗原（抗體）夾心兩步法 Assay processing for One step 25 Assay processing for Two step 18-4 STAT assay proces

10、sing for One step 11 STAT assay processing for Two step 4-4Assay processing for Two step 18-4 (iSystem) 1. At position 1 the sample pipettor dispenses the sample into the RV (reaction vessel). 2. At position 2 the R1 pipettor dispenses the microparticles. 3. At position 3 the vortexer mixes the samp

11、le and microparticles. 4. At positions 4 - 63 the reaction mixture incubates for 18 minutes. 共112個RV杯空位，每18秒轉一個空位，200T/h。常規(guī)28Min/TESTAssay processing for Two step 18-4 (iSystem)5. At positions 64 - 67 the wash zone 1 manifold washes the reaction mixture in the RV, and then removes unbound materials

12、Assay processing for Two step 18-4 (iSystem)6. At position 71 the R2 pipettor dispenses acridinium-labeled conjugate. 7. At position 72 the vortexer mixes the reaction mixture. 8. At positions 73 - 86 the reaction mixture incubates for 4 minutes.Assay processing for Two step 18-4 (iSystem)9. At posi

13、tions 87 - 90 the wash zone 2 manifold washes the reaction mixture in the RV, and then removes unbound materials. Assay processing for Two step 18-4 (iSystem)10. At position 94 the pre-trigger nozzle dispenses Pre-Trigger Solution to the reaction mixture, and then mixes using the vortexer. 11. At po

14、sition 98 the CMIA optical 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. 12. At position 100 the liquid waste arm aspirates the liquid waste from the RV. 13. At position 109 the RV unlo

15、ader removes the RVAssay processing for One step 25 (iSystem) 1. At position 1 the sample pipettor dispenses the sample into the RV (reaction vessel). 2. At position 2 the R1 pipettor dispenses the microparticles and acridinium-labeled conjugate. NOTE: For a delayed one-step assay the R2 pipettor ad

16、ds the acridinium-labeled conjugate at position 71 and the vortexer mixes the reaction mixture at position 72.3. At position 3 the vortexer mixes the sample, microparticles, and conjugate. 4. At positions 4 - 86 the reaction mixture incubates for 25 minutes. Assay processing for One step 25 (iSystem

17、)5. At positions 87 - 90 the wash zone 2 manifold washes the reaction mixture in the RV, and then removes unbound materials. Next position is same to the two step18-4.STAT assay processing for One step 11 (iSystem) 1. At position 47 the STAT pipettor dispenses the sample into the RV (reaction vessel

18、). 2. At position 48 the R2 pipettor dispenses the microparticles and acridinium-labeled conjugate. 3. At position 49 the vortexer mixes the sample, microparticles, and conjugate. 4. At positions 50 - 86 the reaction mixture incubates for 11 minutes. STAT assay processing for One step 11 (iSystem)5.

19、 At positions 87 - 90 the wash zone 2 manifold washes the reaction mixture in the RV, and then removes unbound materials. Next position is same to the two step18-4.STAT assay processing for Two step 4-4 (iSystem) 1. At position 47 the STAT pipettor dispenses the sample into the RV (reaction vessel). 2. At position 48 the R2 pipettor dispenses the microparticles. 3. At position 49 the vortexer mixes the sample and microparticles. 4. At positions 50 - 63 the reaction mixture incubates for 4 minutes. STAT assay processing for Two step 4-4 (iSystem)5. At positions 64 - 67 the wash