ARCHITECT i2000 测试原理 英文手册

i System principles of operationARCHITECT® i System principles of operation provides an overview ofthe CMIA (chemiluminescent microparticle immunoassay)technology, assay processing, and optical system used for analytemeasurement.i System principles of operation topics include:• CMIA method, page 25• Assay processing (i System), page 30CMIA methodCMIA (chemiluminescent microparticle immunoassay) is a detectionmethod used by an i System to measure and quantify analyteconcentration.CMIA method topics include:• CMIA technology and reaction sequence, page 25• Optical measurements (i System), page 28CMIA technology and reaction sequenceCMIA (chemiluminescent microparticle immunoassay) is a technologyused to determine the presence of antigens, antibodies, and analytesin samples.The reactants necessary for CMIA technology include:• Paramagnetic microparticles coated with a capture molecule(antigen, antibody, or viral particle) specific for the analyte beingmeasured• Acridinium-labeled conjugate• Pre-Trigger Solution and Trigger SolutionThe following graphic symbols are used to represent these reactants.Principles of operationi System principles of operation Section 3Section 3-26 Abbott ARCHIT E CT® System Operations Manual96211-102–December, 2003Figure 3.11: Graphical symbols1. Anti-analyte microparticle with capture molecule2. Sample analyte measured3. Acridinium-labeled conjugate4. Sample analyte not measuredA CMIA reaction sequence is the order of interactions between theanalyte present in the sample and the reactants. A sequence is specificto the assay protocol.The following two-step reaction sequence illustrates the basicprinciples of a reaction.1. The R1 pipettor dispenses microparticles (paramagneticmicroparticles coated with capture molecules) into the sample in the reaction vessel. The vortexer mixes the reaction mixture. Figure 3.12: Sample and microparticle binding2. The reaction mixture incubates and the analyte present in the sample binds to the corresponding capture molecules on the microparticles forming the immune complex.3. A magnet attracts the paramagnetic microparticles (bound to the specific analyte) to a wall of the reaction vessel. The wash zone 1 manifold washes the reaction mixture to remove unbound materials. Further processing can now take place.1. Anti-analyte microparticle withcapture molecule2. Sample analyte measured3. Acridinium-labeled conjugate4. Sample analyte not measured Abbott ARCHITE CT® System Operations Manual Section 3-2796211-102–December, 2003Principles of operationSection 3 i System principles of operationFigure 3.13: Magnet attracting paramagnetic microparticles4. The R2 pipettor dispenses a chemiluminescent acridiniumlabeled conjugate. The conjugate binds to the immune complexto complete the reaction mixture.Figure 3.14: Addition of the acridinium-labeled conjugate5. The reaction mixture incubates.6. The wash zone 2 manifold washes the reaction mixture toremove unbound materials.7. The pre-trigger nozzle dispenses Pre-Trigger Solution (hydrogen peroxide) and the CMIA optical system takes a background read.Pre-trigger performs the following functions:– Creates an acidic environment to prevent early release ofenergy (light emission).– Helps to keep microparticles from clumping.– Splits acridinium dye off the conjugate bound to themicroparticle complex. This action prepares the acridiniumdye for the next step.Principles of operationi System principles of operation Section 3Section 3-28 Abbott ARCHIT E CT® System Operations Manual96211-102–December, 20038. The trigger nozzle dispenses Trigger Solution (sodium hydroxide) to the reaction mixture. The acridinium undergoes an oxidative reaction when exposed to peroxide and an alkaline solution. This reaction causes the chemiluminescent reaction to occur.N-methylacridone forms and releases energy (light emission) as it returns to its ground state.9. The CMIA optical system measures the chemiluminescent emission (activated read) over a predefined time period to quantitate the analyte concentration or to determine qualitative interpretations for index (cutoff) assays.Related information...• Assay processing for One step 25 (i System), page 32• Assay processing for Two step 18-4 (i System), page 34• STAT assay processing for One step 11 (i System), page 39• STAT assay processing for Two step 4-4 (i System), page 41 Optical measurements (i System)Optical measurement is the process an i System uses to obtain RLU (relative light unit) readings, and then convert them to assay-specific analyte concentration units or qualitative interpretations for index (cutoff) assays.Optical measurement topics include:• Optical system and measurement sequence (i System), page 28• Data reduction calculation (i System), page 30Optical system and measurement sequence (i System) The optical system on the processing module is a system that directs the chemiluminescent emission from the reaction vessel to the CMIA (chemiluminescent micropartical immunoassay) reader.Figure 3.15: Optical system1. Photomultiplier tube (PMT)2. CMIA reader3. Light pipe4. Trigger Solution delivery nozzle5. Reaction vessel6. Magnet7. CMIA shutter assemblyNOTE: This solution initiates the chemiluminescent reactionthat results in the emission of photons of light.NOTE: The chemiluminescent light produced during this reaction is directly or indirectly proportional to the amount of analyte present in the sample, depending on the type of assay. Measurement occurs as the optical system performs the following: 1. Closes the shutter around the reaction vessel to seal off ambient light2. Turns on the high voltage to the PMT (photomultiplier tube), takes a background read (Pre-Trigger Solution has already been dispensed), and transfers the data to the CPU (central processing unit)3. Dispenses Trigger Solution into the reaction vessel4. Uses the light pipe to collect the emitted light and directs it to the PMT, which is in the CMIA (chemiluminescent microparticle immunoassay) reader5. Takes the activated read by collecting the emitted photons of light6. Transfers the count data to the CPU1. Photomultiplier tube (PMT)2. CMIA reader3. Light pipe4. Trigger Solution delivery nozzle5. Reaction vessel6. Magnet7. CMIA shutter assemblyNOTE: This solution initiates the chemiluminescent reactionthat results in the emission of photons of light.NOTE: The chemiluminescent light produced during this reaction is directly or indirectly proportional to the amount of analyte present in the sample, depending on the type of assay. Principles of operationi System principles of operation Section 3Section 3-30 Abbott ARCHIT E CT® System Operations Manual96211-102–December, 20037. Sums the signal over a defined time period to yield the RLU (relative light unit)8. Turns off the high voltage PMT9. Opens the shutterData reduction calculation (i System)Data reduction calculation is the method used to calculate the final read in RLUs (relative light units). The calculation is:Final Read (RLU) = Activated Read – BackgroundIn performing the data reduction calculation the system:1. Sums the signal measured by the CMIA optical system2. Verifies that:– Background counts fall within an acceptable range– Activated read profile falls within an acceptable set of ranges 3. Subtracts the background counts from the activated read counts to calculate the final read and converted it to concentration units Related information...• i System data reduction methods, page Appendix C-15。