Sysmex® CA-500 Series System Calibration Online Training

Welcome to the Sysmex® 500 Series Calibration online training course. Test methods run on the analyzer need to be calibrated at set intervals, if you are using a new reagent lot number, or if you are directed to do so by the Technical Assistance Center.  NOTE: This course does not include audio. Upon successful completion of this course, you will be able to: Describe basic calibration concepts and tasks necessary to run a calibration Describe why some test methods use a standard curve to calculate final test results and some do not Identify data and key features on the Standard Curve screen Demonstrate how to perform auto dilution calibrations Manually enter MNPT and ISI values for PT calibrations Congratulations. You have completed the Sysmex CA-500 Series Calibration online training course. Listed below are the key points that have been presented. Take time to review the material before you proceed to the final quiz. Describe basic calibration concepts and the tasks necessary to run a calibration. The Sysmex® CA-500 coagulation analyzer uses a standard curve to calculate patient and QC results for most of the tests run on the analyzer. To create a standard curve, the analyzer performs calibrations by running calibrators of known concentration or percent activity and then plotting the change in optical density (Δ OD) or raw clotting time measurements against known calibrator values. You can view calibration data using the Standard Curve screen. Describe why some test methods use a standard curve to calculate final test results, and some do not. For fibrinogen, D-Dimer, and other assays that report final analysis results in concentration or percent activity, the system uses the standard curve to calculate patient and QC results. Some test methods, such as PT, APTT, and thrombin time, do not use a standard curve to calculate final results for patient and QC samples. Instead, these tests report the raw clotting time measurement as the final analysis result—or the tests use the raw analysis result in another way. For example, PT tests use the raw clotting time measurement and MNPT and ISI values to calculate and report out the International Normalized Ratio (INR) and PT Ratio. Identify data and key features on the Standard Curve screen. Note the following calibration data available on the Standard Curve screen. Cal Date: The date the current standard curve was adopted. Reagent names, lot numbers, and expiration dates: Review reagent data before running a calibration to ensure that you are using the same reagents that the system uses to calculate final test results! Lot No. Entry key: Press this key to enter reagent lot number and expiration date. Manual Entry key: Press this key to enter MNPT and ISI values for PT calibrations. Dilution assay values: The system calculates assigned values for each dilution ratio using the initial calibrator value and then plots each value on the X axis of the standard curve. Raw analysis data: The system determines analysis results of each dilution ratio and then plots each result on the Y axis of standard curve. Graph or Print keys: Press these keys to view or print the standard curve for the current calibration. Standard Analysis key: Press this key to open a screen where you can verify or set the calibration analysis parameters and then run an auto dilution calibration. Demonstrate how to perform auto dilution calibrations. From the Main Menu screen, press Standard Curve On the Standard Curve screen, press Select Test, and then select the test key for the calibration data you want to view. On the Standard Curve screen, press Standard Analysis to verify or set the following calibration analysis parameters: Calibrator assay value Dilution set Replicates per dilution Press Start to start the calibration, and then press Graph to evaluate the new standard curve. Press Return, and then do either of the following: Press Set to adopt the new curve. Press Quit to reject the new curve. Manually enter MNPT and ISI values to calibrate PT tests. From the Main Menu screen, press Standard Curve On the Standard Curve screen, press Select Test and then select the PT test key. From the Standard Curve screen for PT, press Manual Entry and then use the keypad to enter MNPT and ISI values. Calibrations are performed by running calibrators of known concentration or percent activity and then plotting the change in optical density (∆OD) or raw clotting time measurements against known calibrator values to create a standard curve. The standard curve is then used to calculate patient and QC results. You can view calibration data using the Standard Curve screen. Not all tests performed on analyzer use a standard curve to calculate patient and QC results. For fibrinogen, D-Dimer and other assays that report final analysis results in concentration or percent activity, the system uses the standard curve to calculate patient and QC results. Check the specific test Application Sheet to see if a standard curve is required. The Sysmex® CA-560 coagulation analyzer is the only instrument in the Sysmex® CA-500 series that can perform the Immunologic test method, D-Dimer. For PT, APTT, thrombin time, and other test methods that report the raw clotting time measurement as the final analysis result, the system does not use a standard curve to calculate results for patient and QC samples. For these reasons, data displayed on a Standard Curve screen vary for different test methods. It depends on the way the system calibrates a test and calculates results. The Standard Curve screen displays reagent information currently used by the system to calculate patient and QC results. During reagent setup, before loading reagents when processing samples, verify that the reagents, lot numbers and expiration dates match the reagent information displayed on the Standard Curve screen to ensure you are using the same reagents that the system uses to calculate final test results The Sysmex® CA-500 series coagulation analyzer calibration Advanced D-Dimer, Fibrinogen, and other assays by creating a standard curve. Use the Standard Curve screen to view current calibration data for a test method or run a new calibration. To open the Standard Curve screen: From the Main Menu screen, press Standard Curve On the Standard Curve screen, press Select Test Select the test for the calibration data you want to view Dilution Set Learn how to define the dilution set for a calibration. Defining the Dilution Set for Calibration Dilution Set No. Dilution Ratios The most common way to create a standard curve is by auto dilution, where the analyzer automatically performs a series of dilutions on a single calibrator. When setting up an auto dilution calibration using the Standard Curve screen, you must define the dilution ratios that you want the analyzer to make. The Sysmex® CA-500 series coagulation analyzer allows you to select from 12 different dilution sets. Each set comprises a series of dilution ratios. Refer to your Application Sheet to determine the appropriate dilution set for the calibration you are running. The table shown here displays the dilution sets available on the Sysmex® CA-500 series coagulation analyzer. 1 1/1, 1/2, 1/4, 1/16/1/32 2 1/2, 4/5, 3/5, 2/5, 1/5 3 1/1 1/2, 1/4, 1/8, 1/16, 0/1 4 1/1, 1/2, 1/3, 1/4, 1/5 5 1/2, 1/4, 1/8, 1/16, 1/31, 0/1 6 10/19, 5/19, 5/38, 1/19, 1/38, 1/95 7 2/1, 1/1, 1/2, 1/4, 1/8, 0/1 8 3/2, 1/1, 1/2, 1/4, 1/8, 0/1 9 2/1, 3/2, 1/1, 1/2, 1/3 10 3/2, 1/1, 1/2, 1/4, 1/8, 1/16 11 4/1, 3/1, 2/1, 1/1, 1/2, 1/3 12 5/4, 1/1, 3/4, 1/2, 1/4 M Set each dilution ratio manually Standard Curve Screen Learn about the Standard Curve Screen. Checklist TitleChecklist TypeChecklist ContentPress Standard Curve KeyHTML From the Main Menu screen, press the Standard Curve key. Press Select Test KeyHTML The test method for the standard curve data is displayed. This screen shows the current standard curve data for the PT test method. To view the standard curve data for the Advance D-Dimer assay, press the Select Test key. Press AdDD KeyHTML Press the AdDD key to view the Standard Curve screen for the Advanced D-Dimer assay. View Standard Curve DataHTML The Advanced D-Dimer Standard Curve screen displays data currently used to calculate AdDD test results (1). The Cal Date (2) indicates when the current standard curve was adopted. Each test method’s Standard Curve screen displays the reagents needed to perform the assay, reagent lot numbers and expiration dates (3), reagent lot numbers used to perform the Advanced D-Dimer calibration (4) (all Advanced D-Dimer assays for patient and QC samples must use these same reagent lot numbers), and reagent expiration dates (5). Press Lot No. Entry KeyHTML Use the Lot No. Entry key to enter or edit reagent lot number and expiration date information. Enter or Edit Regent InfoHTML The system uses the data in these columns to create the standard curve. The left displays the assay value of each dilution made using the Advanced D-Dimer Standard Plasma calibrator. The right column displays the raw analysis data determined for each dilution. To create the standard curve, the system plots the assigned value on the horizontal X axis versus the raw analysis data on the vertical Y axis. Slope is the calculation obtained from a calibration graph where test results for calibrator levels on the Y axis are plotted against expected values for those calibrators on the X axis.   Press Graph KeyHTML To view the standard curve for the current calibration, use the Graph key. Press Print KeyHTML Use the Print key to print the standard curve information and the graph. Select Param KeyHTML  The Select Param key is used by the Technical Applications Specialist (TAS) to define any necessary calculations required to obtain final analysis results. Auto Dilution Calibration Learn how to run an auto dilution calibration. Checklist TitleChecklist TypeChecklist ContentPress Standard Analysis KeyHTML To start an auto dilution calibration, press the Standard Analysis key. Set Initial Calibrator ValueHTML Use the keypad to set the initial calibrator value that the system uses to calculate assay values for each dilution. Use the Up and Down keys to navigate to the Calibrator Value field, if necessary. For example, this screen shows that the initial calibrator value of the Advanced D-Dimer Standard Plasma calibrator is 38.10 mg/L. Define Dilution RatiosHTML Press the Select Dil. Set key until the appropriate dilution set number appears. This screen shows that dilution set selected is 6, which means that the instrument will make the following auto dilutions: 10/19, 5/19, 5/38, 1/19, 1/38, 1/95. The assigned calibrator value for the first dilution ratio is 10/19 x 38.10 mg/L = 20.05 mg/L. Refer to each test method's Application Sheet to determine the appropriate dilution set for the calibration you are running. Set DuplicatesHTML You can also use the keypad to set the number of replicates that you want the instrument to make as it analyzes each dilution for the calibration you are running. Press Start KeyHTML Press the Start key to start the calibration. Select ActionHTML A screen displays asking which reaction tube you want the system to use to process the first dilution. Press Continue to continue using reaction tubes in the order that it last left off from a previous analysis, or press First Tube to start over and use the first reaction tube in the upper-right corner of the reaction tube rack. In this example, First Tube is selected. Message DisplaysHTML The message STD Analyzing. Please wait displays while the calibration analysis takes place. Select Graph KeyHTML Raw analysis data for each dilution ration displays when the calibration analysis is complete. Select the Graph key to evaluate the new standard curve. Evaluate New Standard CurveHTML On the standard curve that appears, the solid line shows the new calibration curve for the Advanced D-Dimer calibration run that has not yet been adopted. When creating standard curves, the system plots the calibrator’s assigned values for each dilution on the horizontal X axis versus the raw analysis data for each dilution on the vertical Y axis. Press the Current Graph key to compare the new curve to the one stored in the system. Compare Standard CurvesHTML Now the graph displays two standard curves. The solid line shows the new standard curve for the Advanced D-Dimer calibration that has not yet been adopted. The dotted line shows the current standard curve for the Advanced D-Dimer calibration stored in the system. Evaluate the new standard curve by comparing it to the current curve and also by running QC. Press the Return key to return to the previous screen. Select ActionHTML Press the Set key to adopt the new standard curve data and return to the Standard Curve screen, or press the Quit key to discard the new standard curve data and return to the Standard Curve screen. In this example, the Set key is select to adopt the new standard curve. Review Standard CurvesHTML The Standard Curve screen appears with the new standard curve data. Your next task will be to run QC to verify that the new Advanced D-Dimer standard curve is valid. Some test methods, such as PT, APTT, and thrombin time, do not use a standard curve to calculate final results for patient and QC samples. Instead these tests either report a raw clotting time measurement as the final analysis result, or the final analysis result is determined another way. For example, for PT tests, the raw clotting time measurement is inserted into an equation to calculate and report out the International Normalized Ratio (INR) and PT Ratio: INR = [PT Clotting time / NMPT]ISI. In this equation: PT Clotting Time = Raw Clotting Time Measurement MNPT = Mean Normal Prothrombin Time ISI = International Sensitivity Index PT Ratio = PT clotting Time / MNPT You must enter MNPT and ISI values on the Standard Curve screen. To find the ISI value, look on the Table of Assay Values packaged with each PT reagent lot. MNPT values are specific for each laboratory. Manual Entry for MNPT and ISI Values Learn how to manually enter NMPT and ISI Values. Checklist TitleChecklist TypeChecklist ContentPress Standard Curve KeyHTML From the Main Menu screen, press the Standard Curve key to open the Standard Curve screen. Press Select Test KeyHTML The Standard Curve screen appears. The test method for the standard curve data displayed appears here. This screen shows that you are viewing current standard curve data for the PT test method. Remember to use the Select Test key to select the test for the calibration data you want to view. View DataHTML The Cal Data (1) indicates when the PT calibration information was last updated. The PT Standard Curve screen does not display calibration data (2) as PT tests do not create a standard curve to calculate final test results. The screen also displays the reagents required to run the test, reagent lot numbers, and expiration dates. Review the Standard Curve screen for each test method as you load reagents to ensure that you are loading the correct reagents. Reagent lot numbers used to perform the PT test appear in this area (3). Remember that all PT tests for patient and QC samples must use these same reagent lot numbers. Reagent expiration dates appear in this area (4). Press Lot No. Entry KeyHTML Use the Lot No. Entry key to enter or edit reagent lot number of expiration date information. Press Manual Entry KeyHTML The MNPT value (used to calculate the INR and PT Ratio) and the ISI value (used to calculate the INR) will have to be entered manually. To manually enter MNPT and ISI values, press the Manual Entry key. Enter NMPT and ISI ValuesHTML A keypad appears that allows you to manually enter the MNPT value in the Normal field, and the ISI value in the ISI field. Remember that the system uses these values to calculate the PT INR. Use the Enter key to enter each value and use the Up and Down arrow keys to move from field to field. Remember that you can find ISI values on the Table of ISI Values packaged with each PT reagent lot. Each lab will need to determine its lab-specific MNPT value. Press the Quit key to close the keypad. Press Set KeyHTML Use the Set key to enter the MNPT and ISI values into the system and return to the Standard Curve screen. Press Print KeyHTML Notice the Graph key is missing as PT tests do not create a standard curve to calculate final test results. Press the Print key to print the Standard curve information. Define CalculationsHTML Your TAS uses the Select Param key to define any necessary calculations required to obtain final analysis results, such as INR and PT Ratio. Load the reagents into their assigned reagent holder positions. Refer to the Application Sheet which lists the reagents, vial types, and reagent holder positions specified for each test method. Next, load the calibrator into the first position in the sample rack.  Auto Dilution Calibration Learn how to run an auto dilution calibration. Checklist TitleChecklist TypeChecklist ContentPress Standard Curve KeyHTML On the Main Menu screen, press the Standard Curve key to open the Standard Curve screen. Press Select Test KeyHTML The Standard Curve screen displays standard curve data used to calculate the PT test. To view the standard curve data for the fibrinogen assay, press the Select Test key. Press the Fbg Test KeyHTML Press the Fbg test key. Press Lot No. Entry KeyHTML The fibrinogen Standard Curve screen appears and displays data that the system currently uses to calculate fibrinogen test results. Before proceeding with the calibration, be sure to press the Lot No. Entry key if you need to enter or edit reagent lot number and expiration date information. Press Standard Analysis KeyHTML Press the Standard Analysis key to begin verifying calibration parameters. Verify Calibrator ValueHTML Use the keypad to enter the fibrinogen assay value of the SHP calibrator in the Calibrator Value field. The system will use this value to calculate the assay value for each dilution. Use the Up and Down keys to navigate to the Calibrator Value field. For this example, the fibrinogen assay value for SHP on the Table of Analytical Values insert sheet is 2.5 g/L. Verify that the correct calibrator assay value appears. Note that it appears in mg/dL units. Verify Dilution Set NumberHTML The dilution set number on the Application Sheet for Fibrinogen is 9. This means that the system will use the SHP calibrator to make 2/1, 3/2, 1/1, 1/2, and 1/3 serial dilutions. Note that the last dilution point has a value of 0.0 mg/dL. There is no need to enter or change any data for this dilution point. If necessary, use the Select Dil. Set key to enter a different dilution set number. Verify ReplicatesHTML The number of replicates for each dilution ratio when performing a fibrinogen calibration is 2. Note that the last dilution point with a value of 0.0 mg/dL has 0 for its replicates number. There is no need to enter or change any data for this dilution point. Now that all the calibration parameters have been verified, press the Start key. Press First TubeHTML If the fibrinogen calibration is the first run of the day after powering on the analyzer, the Continue key will not appear. Press the First Tube key to start the calibration using the first reaction tube in the upper-right corner of the reaction tube rack. The analyzer performs a rinsing program to replace the rinse solution in the hydraulic lines. This takes approximate three minutes. An on-screen message will display during this time. Press Graph KeyHTML During calibration analysis, the following message displays: STD Analyzing. Please wait. As the calibration progresses, raw analysis data appears for each dilution ratio. When analysis is complete, the system calculates the standard curve by plotting the dilution assay values versus the raw analysis data for each dilution. Press the Graph key to evaluate the new fibrinogen standard curve. Press Current Graph KeyHTML Press the Current Graph key to evaluate new and current standard curves simultaneously. The solid line shows the new calibration curve. The dotted line shows the current fibrinogen standard curve stored in the system. Press the Return key to return to the previous screen. Press Set KeyHTML Press the Set key to adopt the new standard curve data and return to the Standard Curve screen. The new calibration data appears on the Standard Curve screen. Perform the following tasks to calibrate test methods that use standard curves to calculate results: Create a new standard curve. The most common way to create a standard curve is by auto dilution where the analyzer performs dilutions on a single calibrator. Evaluate the new standard curve by comparing it to the current standard curve, setting the new calibration data into the system, and printing the calibration data for future use. Run QC to validate the new standard curve. Note: You can also enter calibration data manually, such as to restore a previous standard curve, or to enter MNPT and ISI values necessary to run the PT tests and report INR values. Performing Calibration Learn when to perform calibration. When should you perform a calibration? As specified in the Application Sheet of your reagent If the reagent lot number changes for a test method Every 6 months—even if the lot number of reagent is not changed If you add a new test method to the analyzer If you receive test method updates or software revisions that require a calibration If you have a problem with QC analysis results If you are directed to run a calibration by the Technical Solutions Center (TSC) Refer to the Application Sheets included with each reagent lot for details about product specifications. Application Sheets Learn about information on the Application Sheets. Application Sheets display the following: Reagents needed to run each test method Quality Control products Calibrator used to run a calibration Onboard position and stability of each reagent Dilution set to define calibrator dilution ratios Replications needed for each dilution ratio Test Protocol settings that define how the instrument runs each test method Your Technical Applications Specialist (TAS) can supply Application Sheets to you. You should have an Application Sheet for each test you run on your Sysmex® CA-500 series coagulation analyzer.