Sysmex® CA-500 Series System Coagulation Curve Evaluation Online Training

Welcome to the Sysmex® CA-500 series Coagulation Curves Evaluation online training course. The coagulation curve is a visual display of the activity that occurs during a clotting assay. All coagulation curves comprise similar components; however, each test analysis yields a characteristic pattern or signature coagulation curve. In addition, a test method’s coagulation curve varies if the analysis yields normal or abnormal test results. Note: This course does not include audio.   Upon successful completion of this course, you will be able to: List test methods that generate coagulation curves Identify key features of the Stored Data Graphic Display screen Identify key coagulation curve error codes Describe how to evaluate and troubleshoot key coagulation curve errors Congratulations. You’ve completed the Sysmex® CA-500 Series Coagulation Curve Evaluation online training course. Listed below are the key learning objectives presented. Coagulation test methods, or clotting assays, are unique because they produce a characteristic pattern or “signature” coagulation curve during the assay as the clot is detected. Clotting assays performed on the Sysmex® CA-500 coagulation analyzer include PT, APTT, Fibrinogen, Factor VII, Factor VIII, Thrombin Time, Batroxobin, and Protein C. The Sysmex® CA-500 series coagulation analyzer uses the percent detection method to determine the coagulation time. A reading is taken at the 50% point between the baseline and the endpoint of the coagulation reaction because this is the area of the coagulation curve that is the most linear. Use the Stored Data Graphic Display screen to view the coagulation curve for a clotting assay. Identify key features of the Stored Data Graphic Display screen. To open the Stored Data Graphic Display screen, from the Main Menu screen, press Stored Data, select a sample, and then press Graph. Key features of the Stored Data Graph Display screen include the coagulation curve to provide a visual display of the coagulation reaction, sample flags to describe the analysis sample, Change Scale key to change the Y axis scale, and Output key to print the coagulation curve or output data to a floppy disk or host computer. Identify key coagulation curve error codes. For normal coagulation curves, the clotting assay creates a sigmoid-shaped coagulation curve. If the system generates a coagulation curve error, the system flags test results with an asterisk (*) on the Stored Data screen, and displays error codes on the Stored Data Graphic Display screen. Key error codes include: ERR2: Slight Coagulation, ERR 4: Analysis Time Over, ERR 8: Coagulation Curve Error, ERR 16: Turbidity Level Over, ERR 32: No Coagulation, ERR 100: Range Over, and ERR 128: Early Reaction Error. Describe how to evaluate and troubleshoot key coagulation curve errors. Review the coagulation curve data to determine if you can report test results, or if you must troubleshoot a problem within the assay as follows: Rule out inhibitors, sampling, and reagent problems. Review the coagulation reaction on the Micro Analysis printout. Re-analyze the sample. If the repeat result is not flagged, report the results. If the error occurs again, report the mean of the repeat and initial results if the curves are acceptable and both results are equivalent at the 50% detection point. Do not report results without numeric values. After a sample analysis, the system displays results on the Stored Data screen. Analysis results are displayed as raw data values (sec or dOD) or calculated values (activity %, mg/dL, mg/L, or IU/mL)—it depends on the test method as well as the detection method. The chart shown here displays the three detection methods used to obtain analysis results for several tests performed by the Sysmex® CA-500 series coagulation analyzer. Coagulation test methods, or clotting assays, are unique because they produce a characteristic pattern or signature coagulation curve during the assay as the clot is detected. The Sysmex® CA-560 coagulation analyzer is the only instrument in the Sysmex® CA-500 series that can perform the immunologic method, D-Dimer. Detection Method Test Method Coagulation PT, APTT, Fibrinogen, Factor VII, Factor VIII, Thrombin Time, Batroxobin, Protein C Chromogenic Anthithrombin III, Heparin, Protein C Immunologic D-Dimer To view coagulation curves From the Main Menu screen, press Stored Data Select a sample, and then press Graph The Stored Data Graphic Display screen appears Normal Coagulation Curves Learn how to identify data on normal coagulation curves. Checklist TitleChecklist TypeChecklist ContentView SampleHTML On the Stored Data screen, use the Up and Down arrow keys to select the sample you want to view. Note that for PT tests, coagulation time is shown in seconds. Select Graph KeyHTML For the selected sample, press the Graph key to view the coagulation curves for the coagulation tests that were run.  View Coagulation CurveHTML The Stored Data Graphic Display screen displays the PT coagulation curve (1). Time is shown on the X axis (2). Light scatter intensity is shown on the Y axis (3). For coagulation curves, slope is the calculation obtained from a coagulation graph where light scatter intensity on the Y axis is plotted against analysis time on the X axis. Change ScaleHTML You can change the Y axis scale by pressing the Change Scale key.  View DetailsHTML A 1/1 dilution ratio (1) was performed on the sample, which means that the sample was run straight without dilution. Coagulation time reading (2) was 12.1 seconds. The time reading was taken at the 50% coagulation detection point (3). This is the 50% value between the baseline and the endpoint of the coagulation. The curve is the most stable and linear at this point. The dH reading (4) is the maximum change in light scatter. Detection channel 1 was used for the analysis (5). The error code (6) is 0, which indicates that no error occurred during the analysis. Abnormal Coagulation Curves Learn how to identify data on abnormal coagulation curves. Checklist TitleChecklist TypeChecklist ContentView ErrorHTML The PT result for this sample = 19.6 seconds. An error occurred during the analysis. The asterisk to the left of the result also indicates that an error has occurred. Output GraphHTML Use the Output key to print the coagulation curve or output data to a host computer. An error occurred during the analysis, generating error code 2.  Select Info KeyHTML Press the Info key to open the Error Detail window to view information about error codes. This screen tells you that error code 2 is a Slight Coagulation error. The system generates a Slight Coagulation error if it detects a weak clot formation. Close Error Detail WindowHTML Press the Return key to close the Error Detail window and return to the Stored Graphic Display screen. Data Graphic Display Symbols Learn about the symbols on the Data Graphic Display Screen. Symbols that could appear when analysis calculation could not be performed ---,- Could not calculate a parameter +++,+ Value exceeded displayable range ///./ Could not calculate the mean Symbols that could appear if analysis calculation could be performed but yield abnormal result (displays to left of result) * Replicates are out of set deviation limits, or an error has occurred ! Result was performed on dilution < Result is less than report limit > Result is greater than report limit Symbols that could appear if analysis calculation could be performed but yield abnormal result (displays to right of result) - Result is less than Mark Limit + Result is greater than Mark Limit x Calculated result has not been obtained You can print a coagulation curve that shows the clotting reaction for a specific test result. To print a coagulation curve From the Main Menu screen, press Stored Data Select a sample, and then press Graph to open the Stored Data Graphic Display screen, where you can view the coagulation curve for the selected sample Press Output, and then press Current On the Select Device screen, press IP Graph This list of error codes will help determine if you can report the abnormal test result associated with the error code, or if you must troubleshoot a problem within the assay.  Click here to view or print a PDF of flowcharts to help troubleshoot the coagulation curve errors presented in this course.    ERR 2 Slight Coagulation The system detected a weak clot formation. ERR 4 Analysis Time Over The analysis was not completed within the normal detection time. ERR 8 Coagulation Curve Error Unexpected curve fluctuations occurred. ERR 16 Turbidity Level Over A turbid or lipemic sample crated a dH value greater than the threshold value at the maximum detection time. ERR 32 No Coagulation The system was unable to detect a coagulation reaction (dH <10), or it detected a weak clot (dH >10). ERR 100 Range Over The clotting time at the 50% point was less than the minimum reportable time, i.e., the analysis result was outside the software-defined analysis range. ERR 128 Early Reaction Error The clotting reaction occurred too early or too slowly. Description Action to Take The system generates a ERR 2: Slight Coagulation (SC) error if the system detects a weak clot formation (dH does not reach threshold value). Initial Results: Numeric results will be flagged by an asterisk (*) on the Stored Data List screen. ERR 2 code appears on Stored Data Graphic Display screen and Micro Analysis printout. Possible Causes: Low fibrinogen, factor deficiencies, inhibitors Anticoagulant contamination Reagent problem Check sample for anticoagulant contamination, hemolysis, lipemia, etc. Verify sample and reagent delivery. Review results on the Micro Analysis printout to check for evidence of weak clot formation. Note: Micro Analysis will not print if dH <10. Re-analyze sample. If the repeat result is not flagged with an asterisk (*), report the result. If the error occurs again, report the mean of the repeat and initial results as determined by your laboratory protocol if the curves are acceptable and both results are equivalent at the 50% detection point. Description Action to Take The system generates a ERR 4: Analysis Over Time error if the analysis is not completed within the normal detection time (system detects sample reaction end angle is greater than permitted angle at pre-determined maximum detection time.) Initial Results: Numeric results do not appear on Stored Data List screen. The system displays *** , * ERR 4 code appears on Stored Data Graphic Display screen and Micro Analysis printout. ​Possible Causes: Coagulopathies or medications cause prolonged clotting times Buffer onboard the instrument cold versus at room temperature Check sample for anticoagulant contamination, hemolysis, lipemia, etc. Verify sample and reagent delivery. Set a longer maximum detection time (within the range of 100 to 600 sec) or run PTX for PT Extended tests, or PTTX for PTT Extended tests. Then re-analyze the sample. If you obtain numeric results without an asterisk (*), report the results. If the error occurs again, the sample may not be capable of forming a firm clot. Follow your laboratory’s alternative protocol. Do not report results without numeric values. To print the Micro Analysis format data Go to [Main Menu]→[Special Menu]→[Settings]→[Auto Val/Out]→[Format] Press the Format key until Analysis is selected, and then press Return and Fix Go to [Main Menu]→[Stored Data. Highlight the data to be viewed using the Up and Down arrows Select [Mark]→[More]→[Output]→[Marked]→[IP Graph] Note the following: Do not request Micro Analysis printouts on a mean result. Remember to change the Auto Val/Out setting back to [No Graph] after review of the data. An arrow mark can be observed at the 50% coagulation detection point indicating the clotting time.  If the analyzer is powered OFF and then ON, you lose the ability to obtain the Micro Analysis format data. Description Action to Take The system generates an ERR 8: Coagulation Curve Error if unexpected curve fluctuations occur, such as a sudden rise or fall in light intensity. The system uses six checking methods to determine dH differences at specified points on the curve.  Initial Results: Numeric results do not appear on Stored Data List screen. The system displays *** , * ERR 8 code appears on Stored Data Graphic Display screen and Micro Analysis printout. Possible Causes: Air bubble in reaction tube Samples with artifact, such as protein strands that remain after freezing and thawing plasma ​RBCs in the plasma, platelet clumps, or microclots Check sample for anticoagulant contamination, hemolysis, lipemia, etc. Verify sample and reagent delivery. Review results on the Micro Analysis printout. Re-analyze sample. If the error occurs again, reprint the Micro Analysis printout and report the mean of the repeat and initial results at the 50% mark from both printouts as determined by your laboratory protocol if the curves are acceptable and both results are equivalent. Do not report results without numeric values. Description Action to Take The system generates an ERR 16: Turbidity Level Over error if the dH is greater than the threshold value at the maximum detection time. Initial Results: Numeric results do not appear on Stored Data List screen. The system displays *** , * ERR 16 code appears on Stored Data Graphic Display screen and Micro Analysis printout. Possible Causes: Turbid or lipemic sample Check sample for anticoagulant contamination, hemolysis, lipemia, etc. Verify sample and reagent delivery. Re-analyze the sample. For Fbg and factor assays, dilute using Owren's Veronal buffer. If the repeat result is not flagged with an asterisk (*), report the result. If the error occurs again, follow your laboratory’s alternative protocol, such as ultracentrifugation to remove lipemia. Do not report results without numeric values. Description Action to Take The system generates an ERR 32: No Coagulation (NC) error if it is unable to detect a coagulation reaction (dH <10), or if it detects a weak clot (dH >10) greater than the threshold value at the maximum detection time. Initial Results: Numeric results do not appear on Stored Data List screen. The system displays *** , * ERR 32 code appears on Stored Data Graphic Display screen and Micro Analysis printout. Possible Causes: Low fibrinogen, factor deficiencies, inhibitors Anticoagulant contamination, hemolysis, lipemia ​Reagent problem Check sample for anticoagulant contamination, hemolysis, lipemia, etc. Verify sample and reagent delivery. Review results on the Micro Analysis printout to check for evidence of weak clot formation. Note: Micro Analysis will not print if dH < 10. Review previous patient history to determine acceptability of patient results. Re-analyze sample.   If the error occurs again, reprint the Micro Analysis printout and report the mean of the repeat and initial results at the 50% mark from both printouts as determined by your laboratory protocol if the curves are acceptable and both results are equivalent. Do not report results without numeric values. Description Action to Take The system generates an ERR 100: Range Over error if the clotting time at the 50% point is less than the minimum reportable time. Initial Results: Numeric results do not appear on Stored Data List screen. The system displays *** , * ERR 100 code appears on Stored Data Graphic Display screen and Micro Analysis printout. Possible Causes: Lack of instrument maintenance Reagent contamination Fbg result less than or greater than laboratory-defined redilution limits  PT or APTT results outside reportable ranges For fibrinogen assays, change the dilution ratio and re-analyze (+Fbg & -Fbg). For other assays, do the following: Verify sample and reagent delivery, and inspect instrument condition. Review results on the Micro Analysis printout to check for clot formation. Re-analyze sample. If the repeat result is not flagged with an asterisk (*), report the result. If the error occurs again, visually inspect reaction tube for clot formation or follow your laboratory's alternative protocol. Do not report results without numeric value. Description Action to Take The system generates an ERR 128: Early Reaction Error (ERE) if a clotting reaction occurs too early or too slowly. The system uses four checking methods to detect EREs, and generates an additional error code for each type of ERE detected: ERE 1, ERE 2, ERE 3, and ERE 4. Initial Results: The system will either flag a numeric result or display *** . * on the Stored Data List screen, depending on the ERE code. The system will display both ERR 128 and individual ERE codes on the Micro Analysis printout. Possible Causes: Abnormal sample ​Reagent problem Check sample for anticoagulant contamination, hemolysis, lipemia, etc. Verify sample and reagent delivery. Review results on the Micro Analysis printout to check for clot formation and determine the type of ERE code. Re-analyze sample. If the repeat result is not flagged with an asterisk (*), report the result, as determined by your laboratory protocol. If the error occurs again, review results on the Micro Analysis printout to check for clot formation and determine the ERE code.  If ERE 1 or 4, report the result if the clotting time result correlates with the coagulation curve, and repeat results match initial results at the 50% detection point. If results do not correlate, redraw sample and/or follow laboratory protocol. If not ERE 1 or 4, redraw sample and/or follow laboratory protocol. Do not report results without numeric values. All clotting assays generate coagulation curves. The Sysmex® CA-500 series coagulation analyzer uses the percentage detection method to determine the coagulation time. The majority of the readings are taken at the 50% point between the baseline and the endpoint of the coagulation reaction because this is the area of the coagulation curve that is the most linear. Use the Stored Data Graphic Display screen to view the coagulation curve for a clotting assay. For normal coagulation curves, the clotting assay creates a sigmoid-shaped coagulation curve, such as the curve shown here. If the system generates a coagulation curve error, the system flags test results with an asterisk (*) on the Stored Data screen, and displays error codes on the Stored Data Graphic Display screen. Review the coagulation curve data to determine if you can report test results, or if you must troubleshoot a problem within the assay. . The Sysmex® CA-500 series coagulation analyzer uses the percentage detection method to obtain clotting time results using bH and dH measurements from the coagulation curve. The system monitors each clotting reaction to determine the bH (0% scattered light intensity) and dH (100% scattered light intensity) until the reaction is completed. The system calculates the clotting time at the 50% detection point—the point between the baseline and the endpoint of the reaction where the amount of light scattered per unit time shows the greatest change. To validate the coagulation curve, the system builds algorithms into its software to check if the coagulation curve generated during each clotting assay meets certain criteria. If certain check-method criteria are not met, the system flags the test result and generates a coagulation curve error.