Innovance Heparin Assay Sysmex CS-2500 and CA-660 Procedure PDF, 3/20/2024

Bon Secours Mercy Health Richmond HealthPartners Laboratories Title: Innovance Heparin Assay Sysmex CS-2500 and CA-660 Number: COAG-0035 Revision: 3.0 Approval Date: 3/28/2024 Approved By: Clifton Bridges Laboratory Director SMH RGH (CLBRIDGE) Review Date:...

Bon Secours Mercy Health Richmond HealthPartners Laboratories Title: Innovance Heparin Assay Sysmex CS-2500 and CA-660 Number: COAG-0035 Revision: 3.0 Approval Date: 3/28/2024 Approved By: Clifton Bridges Laboratory Director SMH RGH (CLBRIDGE) Review Date: 3/20/2024 Reviewed By: CoraGabriela UramTuculescu Laboratory Director (URAM0000) COAG-0035 Innovance Heparin Assay Sysmex CS-2500 and CA-660 Copy of version 3.0 (approved and current) Last Approval or Uncontrolled Copy printed on 12/12/2024 11:22 3/20/2024 Periodic Review Completed AM Next Periodic Review Leetta Bateman Lead MLS 3/20/2025 Printed By Needed On or Before (lcbatema) Effective Date 3/20/2024 Organization Richmond Market Approval and Periodic Review Signatures y p M co11:22 Type Description Date Version Performed By Notes Approval Laboratory Director 3/28/2024 3.0 Clifton Bridges Laboratory Director SMH RGH (CLBRIDGE) A Approval Laboratory Director 3/21/2024 3.0 d Jack Wilson Laboratory Director SPEC RCH (JJWILSON) e 24 l l 0 Approval Lab Director 3/20/2024 3.0 t r 12/1o 2/ 2 CoraGabriela UramTuculescu Laboratory Director (URAM0000) Approval Laboratory Director 3/18/2024 3.0 o n f Iris Martin Laboratory Director (MART0216) o c ta RCH s n rren Approval Laboratory Director SECC 3/14/2024 3.0 U Cu Angie DUONG Laboratory Director (ADUONG) Laboratory Approval Director 3/14/2024 3.0 Jeffery Green MD (JTGREEN) SFMC WEC Owner Approval 3/14/2024 3.0 Approval Thuy Sandy Pham Education Coordinator (101706) Laboratory Approval Director 3/14/2024 3.0 MRMC CoraGabriela UramTuculescu Laboratory Director (URAM0000)  New Lab Approval Director 3/6/2024 2.0 Approval CoraGabriela UramTuculescu Laboratory Director (URAM0000) Periodic Designated 11/7/2023 2.0 review Reviewer Thuy Sandy Pham Education Coordinator (101706) New Lab Approval Director 9/14/2023 2.0 Jeffery Green MD (JTGREEN) Approval Recorded on 5/5/2023 by Thuy Sandy Minakowski Education Periodic Coordinator (101706) review Designated when document Captured 12/20/2022 2.0 Thuy Sandy Pham Education Coordinator Reviewer added to Document outside Control (previous MediaLab system of record: Master Control Document Records) y p M Recorded on 5/5/2023 by Thuy Sandy Approval Captured outside Lab Director 12/9/2022 2.0 Clifton Bridges Laboratory Director (CLBRIDGE) o c :2 2 A Minakowski Education Coordinator (101706) when document added to Document MediaLab d e 24 11 Control (previous system of record: l l 0 Master Control Document Records) t r 12/1 o 2/ 2 Approvals and periodic reviews that occurred before this document was added to Document Control may not be listed. Version History Version Status Type o n f o Date Added Date Effective Date Retired 3.0 2.0 Approved and Current Retired Major revision c t as n rren First version in Document Control 3/13/2024 5/5/2023 3/20/2024 12/20/2022 Indefinite 3/20/2024 U Cu  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 CLINICAL SIGNIFICANCE Heparin is a commonly used anticoagulant for the prevention and treatment of thromboses and embolisms. Heparin works as an anticoagulant by inactivating thrombin and coagulation factor Xa through binding antithrombin (AT). Due to this inactivating mechanism, the Heparin assay is also commonly referred to as an anti-Xa assay. Heparin exists in two forms: unfractionated heparin (UFH) and low-molecular-weight-heparin (LMWH). UFH and LMWH preparations are widely used as prophylactic and therapeutic anticoagulants. The main clinical indications for UFH are in prevention and treatment of venous thromboembolism, in certain types of coronary artery syndrome and in thrombotic stroke. LMWH has been replacing UFH in many of the latter’s traditional indications. Due to numerous y p M patient. o influences, the anticoagulant effect of an identical dose of heparin varies from patient to c :2 2 A d e 24 11 PRINCIPLE l 20 l complexity for the quantitation of heparin (UFH and LMWH) t / r 1 2 The Innovance Heparin assay is an automated, one-stage chromogenic /o assay of moderate 2 activity in the plasma of patients undergoing heparin anticoagulation therapy. The Innovance o o 1n f Heparin assay is performed on the n c Sysmex CS-2500 and CA-660 coagulation systems. The performance of this assay has not been as established in neonate and pediatric patient populations. t nreagents. The reagent kit consists of a Reagent e component, containing Xa, and a Substrate specific for Xa. When the componentsC U The Heparin assay utilizes ready-to-use liquid r ur component, containing a chromogenic substrate are mixed, Xa converts the chromogenic substrate into two products, one of them being paranitroaniline. The formation of paranitroaniline is then quantified by the coagulation analyzer employing light absorption at a specific wavelength of 405 nm. In the presence of a heparin-containing sample, the formation of paranitroaniline will be reduced in a time dependent manner. This is due to inhibition of Xa by the heparin/AT complex. This complex is formed in the patient's plasma and competes with the substrate conversion by Xa. The concentration of the complex is not only dependent on the concentration of heparin, but also on the availability of the patient’s endogenous antithrombin. When compared to the reference curve, it is possible to quantify the activity of heparin in a patient’s plasma, allowing for the verification of intended anticoagulant target levels. Page 1 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 1 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 SPECIMEN Patient Preparation: No special patient preparation is required. Specimen Type Platelet-poor, Sodium Citrate plasma. Patients with HCT >55% require adjusted anticoagulant tubes. See Calculations section for instructions on adjusting sodium citrate tubes for blood collection. Specimen Volume Properly filled 3.2% Sodium Citrate tubes (a minimum fill of 90%). Collect venous blood according to manufacturer’s instructions. Mix 1 part 3.2% sodium y p M citrate solution with 9 parts venous blood, avoiding the formation of foam. An evacuated tube system or a syringe may be used. o c :2 2 A Handling Conditions d e 24 11 l l Check whole blood specimen for any clot formation by gentle inversion and observation. Centrifuge the specimen as soon as possible after collection for 15 minutes at0 t r 12/1o 2/ approximately 2000 x g to produce platelet-poor plasma (PLT 4 hours old, unless received frozen. y p M during transport). o Frozen sampled received thawed in the laboratory (frozen samples must remain frozen c :2 2 A EQUIPMENT, MATERIALS, AND REAGENTS d e 24 11 l l 0 Analyzer t r 12/1o 4 mL sample cups 2/ 2 Equipment and Materials o n f Reaction tubes o SLD mini cups Sysmex CS-2500 Coagulation c ta n rren s GW 5 reagent caps GW 5 compatible cover rings Analyzer U Cu Barcoded reagent and control racks Barcoded sample racks Waste container Deionized water 2 mL or 4 mL sample cups Reaction tubes Sysmex CA-660 Coagulation Barcoded sample racks Analyzer Waste container Rinse bottle Deionized water Page 3 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 3 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Reagent Storage and Stability Opened Stability CS-2500 CA-660 Reagent Unopened Stability (when stored in On-board On-board closed, original vial) Stability Stability Innovance Heparin Until printed expiration 8 weeks at 2-8°C 72 hours 16 hours Reagent Kit when stored at 2-8°C Owren’s Veronal Until printed expiration 8 weeks at 2-8°C 8 hours 8 hours Buffer (OVB) when stored at 2-8°C Until printed expiration CA CLEAN I when stored at 2-8°C 30 days at 2-8°C y 120 hours p M 24 hours Reagent Preparation o c :2 2 Heparin reagents (Reagent and Substrate) are liquid and ready to use. Before use, A d 11 unscrew and remove caps. Note: Do not pour reagents from one bottle to another. e 24 other lots may lead to incorrect results. l l o All kit components are lot specific. The combination with components from 0 t 1. On the instrument screen, select the Reagent2r o / 2 CS-2500: reagents are placed into a reagent rack, then2placed on the reagent table. /1icon > highlight reagent position > empty/expired reagents. o o n select Change/Add > lift reagent sectionf 1 lid, cover, and lift out rack > remove n cas showing > load rack, replace cover and lid. 2. Add new vial to rack with 1D barcode Select OK > barcode reading will t n occur. e U 3. On Reagent screen, touchuthe Change to update date C rr reagent position that was just loaded > press and time. CA-660: reagents are placed directly onto cold reagent positions of the reagent table. o Note: Heparin and D-dimer assays cannot be performed at the same time on the CA series analyzer as both require the designated cold reagent spots. CALIBRATION Calibration Material INNOVANCE® Heparin Calibrator (lyophilized material) o Level 1: contains no heparin o Levels 2-5: contains defined levels of UFH and LMWH Page 4 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 4 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Calibration Frequency Every 6 months. With every new lot of Innovance Heparin. After major maintenance or service, if indicated by quality control. As indicated by laboratory quality control procedures. Calibrator Storage and Stability INNOVANCE® Heparin Calibration Material Reconstituted Stability Unopened Stability (When stored in closed, original vial) CS-2500 On-board Stability y p M CA-660 On-board Stability Until stated expiration when stored at 2-8°C 24 hours at 15-25°C 48 hours at 2-8°C 8 hours o c :2 2 A 4 hours Calibration Preparation d e 24 11 1. Reconstitute Heparin calibrator with 1.0 mL of deionized water. l l 0 2. Allow vials to stand for at least 15 minutes at 15-25°C following reconstitution. 3. Mix vials gently before use. Calibration Procedure t r 12/1o 2/ 2 n f Refer to the Operator’s Manual and Reference Guide for further details on calibrations. o o Pipetting, mixing, and processing of calibrator levels is performed automatically by the analyzer. c ta n rren s A single, universal calibration curve is generated for both UFH and LMHW. CS-2500 Calibration: U Cu 1. Transfer calibrators into SLD mini cup and replace in corresponding GW5 vials > place vials in yellow C-rack > load rack onto analyzer. 2. On Reagent screen, update newly loaded calibrators. 3. From Order screen > select Switch Order > Holder Calib Curve Order > select Heparin > Change > OK > select the correct calibrator lot from list > OK > Start. 4. Review calibration status and progress in Joblist. 5. Compare new calibration curve to the current calibration curve prior to validating the calibration. CA-660 Calibration: 1. Before starting new calibration, print the old curve. 2. Load reagents for calibration. Page 5 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 5 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 ▪ Transfer calibrators into 2 mL or 4 mL sample cups and place in 4 mL sample rack. ▪ Replenish CA Clean I and load fresh, room temperature OVB. 3. Change Test Group to view test to be calibrated. 4. From Main Menu, select Standard Curve > Select Test > select parameter > verify lot numbers. 5. Select Standard Analysis > enter calibrator value in cursor. ▪ Scan 2D barcode > press Register. 6. Accept calibration > Set > Print. Perform QC. QUALITY CONTROL y p M Quality Control Material INNOVANCE Heparin UFH Control level 1 o c :2 2 A INNOVANCE Heparin UFH Control level 2 INNOVANCE Heparin LMWH Control level 1 d e 24 11 INNOVANCE Heparin LMWH Control level 2 l l 0 QC Frequency t r 12/1o 2/ 2 Two sets of two levels of QC are performed: 1. Every 8 hours of patient testing. o n f o 2. When reagent vial is changed. 3. Following a calibration. c ta n rren s 4. If there is reason to question the accuracy or precision of the test system. All QC must be: U Cu Performed in the same manner as a patient sample. Found acceptable prior to resuming patient testing. Documented in Unity, including any corrective actions. QC Storage and Stability INNOVANCE® Heparin UFH and LMWH Controls CS-2500 CA-660 Unopened Stability Opened Stability On-board Stability On-board Stability 24 hours at 15-25°C Until stated expiration 48 hours at 2-8°C 8 hours n/a when stored at 2-8°C 4 weeks at -20°C Page 6 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 6 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 CS-2500 QC Procedure: Refer to the Operator’s Manual and Reference Guide for further details on performing QC. 1. Transfer controls into SLD mini cup and replace into the vial > place vial in the yellow C- rack. 2. From Order screen > Switch Order > Holder QC Order > Order Entry > select QC01- QC20 tab > QC file > select Heparin assay > OK > Start. ▪ Refer to the quick reference guide to process controls from the reagent table or sample rack. CA-660 QC Procedure: Refer to the Operator’s Manual and Reference Guide for further details on performing QC. y p M 1. Transfer QC material into sample cups (2 or 4 mL). o c :2 2. From the Main Menu > press Test Group > select the test group that includes 2 A Heparin > press FIX to return. d e 24 11 3. Select No. ID Entry > Press QC and the file number > select tests to run > Enter > Quit. l l 0 4. Place rack into sampler > select Start. 5. Review and print QC. t r 12/1o 2/ 2 PROCEDURE o n f o c ta s Pipetting of samples and reagents, as well as mixing and processing, is performed automatically n rren by the analyzer. Refer to the Quick Guide or Operator’s Manual for additional details. Patient Testing on CS-2500 Assay name on analyzer: INN-Hep U Cu Automated Sample Processing: 1. Ensure Host Connection status is green or orange. 2. Place sample into barcoded sample rack. 3. Place rack on the sampler and select Start. 4. Confirm sample order status on the Joblist. Manual Sample Processing: 1. Place sample into barcoded sample rack. 2. Place rack on the sampler. 3. On Order screen, enter rack number and sample ID in corresponding tube position. 4. Select assay to be performed >OK > Start. 5. Confirm sample order status on the Joblist. Page 7 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 7 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Micro Mode Sample: 1. This mode is selected for low volume samples. Patient Testing on CA-660 Assay name on analyzer: IHep Automated Sample Processing: 1. Set samples with barcode facing away from analyzer. 2. From the Main Menu > press Test Group > select the test group that includes Heparin > press FIX to return > press Start. 3. Select Tube Position. ▪ First Tube = start at first sample reaction tube position y p M Manual Sample Processing: o c :2 2 A ▪ Continue = start at next sample reaction tube position after the last run d 11 1. From the Main Menu, press Test Group > select the test group that includes Heparin > press FIX to return. e 24 2. From Main Menu, press ID No. Entry and enter sample ID. l l 0 o 3. Select parameter to be performed > press Enter > press Start. ▪ O = ordered t r 12/1 2/ 2 ▪ - = not ordered 4. Select Tube Position. o n f o c ta s ▪ First Tube = start at first sample reaction tube position n rren ▪ Continue = start at next sample reaction tube position after the last run U Cu Once a sample is loaded, the Heparin assay takes two minutes to complete each run. CALCULATIONS Patients with a HCT >55% must be drawn with an adjusted amount of sodium citrate. Samples with a HCT >55% are identified visually by the tech after centrifugation and may not produce an instrument error code. This value may be confirmed by review of patient laboratory CBC most recently reported, as well. This adjusted collection must be coordinated by the laboratory. The corrected ratio of blood to anticoagulant is calculated using the following formulas: For samples drawn in 4.5 mL glass or plastic tubes, the correction formula is as follows: 𝐶 = 1.85 𝑥 0.001 (100 – 𝑃𝑎𝑡𝑖𝑒𝑛𝑡’𝑠 𝐻𝐶𝑇) 𝑥 4.5 Page 8 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 8 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Example: Polycythemic patient has a HCT of 60%. C = 1.85 x 0.001 (100-60) x 4.5 C = 1.85 x 0.001 (40) (4.5) C= 0.333 mL = 0.33 mL of sodium citrate needed. The BD 4.5 mL glass citrate tubes contain 0.5 mL of sodium citrate: 0.5 – 0.33 = 0.17 mL of sodium citrate removed from the 4.5 mL tube. For samples drawn in a 2.7 mL plastic tube, the correction formula is as follows: y p M Example: 𝐶 = 1.85 𝑥 0.001 (100 – 𝑃𝑎𝑡𝑖𝑒𝑛𝑡’𝑠 𝐻𝐶𝑇) 𝑥 2.7 o c :2 2 A Polycythemic patient has a HCT of 60%. d e 24 11 C = 1.85 x 0.001 (100-60) x 2.7 l l 0 C = 1.85 x 0.001 (40) (2.7) t r 12/1 C = 0.1998 = 0.20 mL of sodium citrate needed. o 2/ 2 o of sodium citrate removed from the 2.7 mL tube. n f The BD 2.7mL plastic citrate tubes contain 0.3 mL of sodium citrate: 0.3 – 0.20 = 0.10 mL o c ta s Important! Be sure to mark the tube with the correct draw volume before providing it to the n rren individual performing phlebotomy. Instruct the collector that the vacuum is broken, and the U Cu tube will need to be filled by syringe (with needle removed) to the indicated mark. DILUTIONS No dilutions are performed for the Heparin assay. REPORTING RESULTS Test Code: HEPXAB Unfractionated and Low Molecular Weight Heparin Reference Range: Heparin is not expected to be present in individuals unless used therapeutically. Page 9 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 9 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Reporting Format: Heparin results are reported in IU/mL. Do not report dOD results. Therapeutic Ranges: UFH: 0.3 – 0.7 IU/mL LMWH: 0.6 – 1.0 IU/mL Analytical Measurement Range (AMR): 0.10 – 1.50 IU/mL Clinical Reportable Range (CRR): 0.10 – 1.50 IU/mL Critical Results: ≥1.0 IU/mL Troubleshooting Results Over Range (> ---.--) y p M 1. Confirm specimen integrity. o c :2 A a. Sample is not clotted, hemolyzed, lipemic or icteric, and tube is sufficiently filled. 2. Compare Heparin results with aPTT results from same tube. Run aPTT offline if not 2 previously ordered. d e 24 11 interference. See drugs listed below. l l a. A normal aPTT result and high Heparin (> ---.-- ) result may indicate medication 0 t r 12/1o 2/ 2 b. A high aPTT result and high Heparin (> ---.-- ) result may indicate Heparin contamination or a truly elevated Heparin result. specimen has been drawn improperly. o on f 3. Rule out contaminated line draw or a draw from above IV line. Request recollection if c ta s a. A draw from the opposite arm of the IV line is preferred. If not possible, nurse n rren should pause heparin infusion for 15 minutes and draw coagulation specimen U Cu peripherally in the same arm, below the heparin infusion. Refer to Pharmacy IV Heparin Infusion Policy for additional details. 4. Confirm patient is not taking any interfering medications. a. Interfering anticoagulants include Apixaban (Eliquis), Danaparoid Sodium (Orgaran), Fondaparinux (Arixtra), and Rivaroxaban (Xarelto). b. Confirmation is obtained by calling the caregiver or RN responsible for the patient. Results Over Range (> ---.--) Action for Heparin Do not report the numeric result. If specimen integrity is questionable: Credit entire accession as INTEG. Do not report the numeric result. If patient is medicated with an Credit HEPXAB as SUA and attach free text comment: anticoagulant listed: “Unable to obtain results due to specimen interference.” Page 10 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 10 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Results Over Range (> ---.--) Action for Heparin Report the result as >1.5 IU/mL. If the above investigation is acceptable: Attach free text comment: “RN confirms that patient is not medicated with an interfering anticoagulant.” Note: Properly document crediting and critical notifications to caregivers with last name, credentials, time, and tech code or initials. See QA policy for further details. Reporting in the LIS: Site MRMC Worksheet MCOAG y p M RCH RGH SFMC CCOAG RCOAG FCOAG o c :2 2 A SMH SCOAG d e 24 11 CAP Surveys l l 0 r 12/1o 2/ 2 When performing proficiency testing, select ‘hybrid curve’ as the calibration curve. t MAINTENANCE o on f c ta s All maintenance is to be recorded in the SmarTerm QCE function. QCE sheets can be located in the QCE folder on the Shared Drive. n rren CS-2500 MAINTENANCE U Cu CS-2500 Maintenance Maintenance Frequency Maintenance Task Instrument shutdown Check and discard trap chamber fluid Remove condensation from reagent table and reagent table cover Daily Instrument start up Discard used cuvettes Discard waste fluids Document daily maintenance Remove water from tray No. 48 Weekly Clean the Main Unit Clean the Rinse Tank Page 11 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 11 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 CS-2500 Maintenance Maintenance Frequency Maintenance Task Replace trash box liner Document weekly maintenance Clean filters Monthly Document monthly maintenance Adjust pressure and vacuum when errors occur Prime hydraulic line Remove cuvette blockages As needed Wipe the piercer Confirm unit and parts cycle y p M Clean Sysmex racks Replace supplies (lamp, fuses) Document as-needed maintenance o c :2 2 A Instrument Shutdown d e 24 11 l l During the daily shutdown, a rinse probe is performed. Verify that CA Clean I is on board, 0 sufficient in volume, and still within stability. 1. Select “Shutdown” on the toolbar. t r 12/1o 2/ 2 2. 3. o n f A dialog box will appear > select “Turn the Main Unit OFF.” Shutdown progress will take approximately 5 minutes. o 4. 5. 6. c ta n rren s Once shutdown is complete, a dialog box will appear > select “OK.” Select Red “X” in upper right-hand corner of screen to exit the IPU > select “OK.” Shutdown Windows by selecting the start menu icon > Shutdown. 7. U Cu Turn the main unit switch off, located on the side of the instrument. a. Do not switch the main unit off if reagent cooling was previously selected. Check and Discard Trap Chamber Fluid 1. Open the pressure adjustment area cover on the left side of the Main Unit 2. Inspect the trap chamber for fluid or condensation. 3. If any fluid is present, remove the trap chamber. 4. Remove the float and dispose of the fluid collected in the chamber. 5. Return the float to the chamber and reinstall the trap chamber. 6. Close the pressure adjustment area cover. Remove Condensation from Reagent Table and Reagent Table Cover 1. Open the reagent table cover. 2. Pull out all reagent racks. 3. Remove condensation from the reagent table and reagent racks with paper towels. Page 12 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 12 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 4. Remove condensation from the reagent table covers. 5. Replace the reagent racks. 6. Replace the reagent table covers and light shield lid. Instrument Start Up 1. Press the power button to turn the IPU power on. 2. Log into Windows using the CS administrator password. 3. Once the system’s database has opened, log into the system. 4. Turn the Main Unit power switch on. a. Once the status display changes from warming up to ready, analysis may begin. Discard Used Cuvettes y p M If the instrument is used continuously, discard the used cuvettes at least once every 24 hours or before the number of discarded cuvettes exceeds 500. 1. Take out the cuvette trash box. o c :2 2 A 2. 3. Discard used cuvettes. Return the cuvette trash box to its original position. d e 24 11 4. l l Reset the trash counter in the software by selecting “Yes” on the Reset Trash Counter 0 dialog box. Discard Waste Fluids t r 12/1o 2/ 2 1. Select “Change Waste Tank” on the Operation Panel of the Maintenance screen. 2. Remove the cap of the rinse tank. 3. Lift the float switch from the tank. o on f 4. Discard the waste fluids. c ta n rren s 5. Attach the float switch to the waste tank and tighten the cap. 6. Check for any kinks in the tube. U Cu 7. Select “OK” on the Change Waste Tank dialog box. Remove Water from Tray No. 48 1. Pull out and remove tray No.48 located beneath the center of the analyzer. 2. Remove any water that has accumulated. 3. Replace tray No.48. Clean the Instrument 1. Weekly, clean the instrument exterior when the instrument is shutdown. a. Wipe the instrument exterior with a paper towel moistened with water and neutral detergent. b. Wipe again using a soft, dry paper towel. 2. Weekly, clean the instrument interior when the instrument is shutdown. a. Open the light shield lid and the reagent section lid. b. Wipe with a paper towel moistened with water and neutral detergent. Page 13 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 13 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 c. Wipe again using a soft, dry paper towel. d. Close the light shield lid and reagent section lid. 3. Weekly, clean the Rinse Tank. a. Remove the cap of the rinse tank and lift the float switch from the tank. b. Rinse the inside of the tank and float switch with tap water or 70% isopropyl alcohol, then rinse with distilled water. c. Replenish the rinse water. d. Attach the float switch to the rinse tank and tighten the cap. e. Check for any kinks in the tubing. Cleaning the Filters 1. Remove the filters. y p M a. Filter No. 16A: Left side of the Main Unit b. Filter No. 513 and Filter No. 514: Back right of the Main Unit 2. Use a vacuum cleaner or similar tool to remove dust. o c :2 2 A 3. Replace the filters. d e 24 11 CA-660 MAINTENANCE l l 0 t r 12/1 CA-660 Maintenance o 2/ 2 Maintenance Frequency o n f Shutdown the instrument o Maintenance Task c ta n rren s Clean the sample probe Check and drain trap chamber Discard used reaction tubes* Daily U Cu Remove condensation from reagent rack Dispose of waste* Check and record temperatures Check and replace as needed: Rinse solution, reaction tubes, CA Clean I, CA Clean II, OVB, and other reagents. Weekly Clean interior and exterior of instrument Clean rinse bottle Quarterly Perform LED calibration** Check consumables: Rinse solution, reaction tubes, reagents Replace fuses As needed Replace printer paper Perform Rinse & Prepare *These tasks may be performed more often than daily, as needed. **LED calibration is required every 30 days for laboratories reporting a derived fibrinogen result. Page 14 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 14 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Instrument Shutdown 1. Turn off power, wait 15 seconds, and turn back on. Clean Sample Probe 1. Clean probe exterior. 2. Verify that cleaners are set and contain sufficient volume. a. CA Clean I in position 11. b. CA Clean II in position 13. 3. Execute Probe Rinse a. On the Main Menu screen, press Special Menu > Rinse Probe. b. Screen will display “Ok to rinse Probe?” > press Set. c. Probe rinse progress will display on the screen, approximately 9 minutes. y p M Drain Trap Chamber 1. Check the pneumatic trap chamber for fluid. Drain if necessary. o c :2 2 A Empty Reaction Tube Trash Drawer d e 24 11 Trash drawer holds approximately 60 used reaction tubes. l l 0 emptied when necessary. Clean drawer with tap water, dry, then replace drawer. t r 12/1o System does not keep track of trash count, drawer must be checked regularly and 2/ 2 Check and Record Temperatures o on f c ta 1. On the Main Menu screen, select Sysmex > Temperature. n rren 2. The temperature of each component is displayed. 3. Record temperatures in Unity. s Replenishing Consumables U Cu During analysis: On the confirmation screen, select “Conf.” When the system Rinse Solution status becomes ready, replenish the rinse solution in the external container. Press Start key to resume analysis. Reaction Tubes Completely refill both reaction tube trays and select “First Tube.” CA Clean I Loaded into position 11 and used for cleaning during sample analysis. Loaded into position 13 and used during maintenance rinse and for specific CA Clean II assays. Use a fresh container every with every replacement in position 12. Used as OVB diluent diluent on specific assays and while performing standard curve calibrations. Do NOT top off existing container. Cleaning the CA-660 Analyzer 1. Clean with 1:2 diluted CA Clean I. Page 15 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 15 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 2. Wipe with a towel moistened with water. 3. Dry with a soft towel. Cleaning the Rinse Bottle 1. Remove the rinse bottle cap by turning it counterclockwise. 2. Empty the contents of the bottle.. 3. Add a sufficient amount of 70% isopropyl alcohol to coat the interior surfaces of the bottle. 4. Shake the bottle to coat the interior surfaces. 5. Allow to sit for 10 minutes and empty. 6. Rinse the bottle with fresh deionized water. 7. Refill the bottle and reassemble. y p M LED Calibration 1. When LED calibration screen appears, select OK. o c :2 2 A 2. d 11 Navigate to the Special Menu > Special Operate > LED Calibration Menu > Calibration. e 24 3. 4. l l Identify and enter the CA CAL S target value from the insert sheet. Select Vial Type by using the arrow keys. 0 5. 6. Load calibrator into position 1. Set CA Clean I into position 11. t r 12/1o 2/ 2 7. 8. o on f Close light shield and select Set to initiate LED calibration. Upon completion, LED Update Confirmation screen appears. c ta a. Press FIX to accept the calibration. n rren b. Press Cancel to reject the calibration. s U Cu As-needed maintenance tasks can be located in the CS-2500 and CA-660 Operator’s Manuals or online at Siemens PEP. See chart above for as-needed maintenance tasks. PROCEDURAL NOTES The Heparin assay is not to be used for the determination of the activity or concentration of anticoagulants other than UFH and LMWH (e.g., heparinoids, synthetic pentasaccharides, direct factor Xa inhibitors). Results should always be interpreted in conjunction with the patient’s medical history, clinical presentation, and other findings. Dispose of hazardous or biologically contaminated materials according to the practices of your institution. Discard all materials in a safe and acceptable manner and in compliance with all government requirements. Page 16 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 16 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 For in-vitro diagnostic use only. The performance of this device has not been established in neonate and pediatric patient populations. To reduce the influence from heparin antagonists, such as platelet factor 4 (PF4), dextran sulfate is included in the reaction mixture. LIMITATIONS OF THE PROCEDURE Sensitivity The limit of quantitation is below the lower limit of the assay range (0.10 IU/mL) and was determined according to CLSI guideline EP17-A25 using UFH and LMWH containing y p M plasmas (Total error of ≤0.085 IU/mL). Limits of detection o c :2 Due to matrix effects, inter-laboratory survey samples and control samples may yield 2 A d 11 results that differ from those obtained with other methods. It may therefore be e 24 l l necessary to assess these results in relation to method-specific target values. 0 In Vivo Interferences t o r 12/1 2/ 2 Turbidity and particles in the samples may interfere with the assay. Therefore, samples o n f containing particles must be centrifuged prior to testing. Lipemic or turbid samples which cannot be clarified by centrifugation (10 minutes at approximately 15,000 x g) o must be excluded from the assay. Endogenous Substances: c ta n rren s Interferent Bilirubin (not conjugated) U Cu CS-2500 No Interference up to… 37 mg/dL CA-660 No Interference up to… 20 mg/dL Bilirubin (conjugated) 39 mg/dL 28 mg/dL Hemoglobin 0.692 g/dL 0.134 g/dL Platelet Factor 4 0.7 µg/mL 0.7 µg/mL Triglycerides 414 mg/dL 267 mg/dL Exogenous Substances: Interferent No Interference up to… Apixaban (Eliquis) 4.1 ng/mL Danaparoid Sodium (Orgaran) 0.03 IU/mL Fondaparinux (Arixtra) 35.5 IU/mL Rivaroxaban (Xarelto) 7.1 ng/mL Page 17 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 17 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 REFERENCES 1. Drucker Diagnostics, PA, USA. HORIZON 6 Flex, 6 Flex FA Operator’s Manual, 03-0-0002- 0138 Rev. C. 2. Hirsh, Jack, Anand, Sonia S., Halperin, Jonathan L., Fuster, Valentin. “Mechanism of Action and Pharmacology of Unfractionated Heparin.” Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 21, p. 1094-1096, 2021. https://doi.org/10.1161/hq0701.093686. 3. Siemens Healthcare Diagnostic Inc., USA. BCS XP System, Sysmex CS-2500 System, Sysmex CS-5100 System, Heparin / INNOVANCE Heparin Technical Procedure, 10873535GU11 Rev. 01 – EN, February 2013. y p M 4. Siemens Healthcare Diagnostic Inc., USA. INNOVANCE Heparin Expanding precision medicine, LD-20-NAM-655, February 2020. o c :2 2 A d 11 5. Siemens Healthcare Diagnostic Inc., USA. Sysmex CS-2500 System Quick Reference e 24 Guide, SMN: 11554034 Rev. T05013.001, May 2021. l l 0 o 2/ 2 6. Siemens Healthcare Diagnostic Products, USA. INNOVANCE Heparin Calibrator Package Insert, 10873530GU11 Rev. 01 – en, January 2017. t r 12/1 o on f 7. Siemens Healthcare Diagnostic Products, USA. INNOVANCE Heparin UF Control 1 and 2 / January 2017. c ta INNOVANCE Heparin LMW Control 1 and 2 Package Insert, 10873534GU11 Rev. 01 – en, n rren s U Cu 8. Siemens Healthcare Diagnostic Products, USA. INNOVANCE Heparin Package Insert, 11541651_en Rev. 04, November 2021. 9. Siemens Healthcare Diagnostic Products, Germany. Sysmex CA-660 series Reference Guide, Application Sheet Heparin / INNOVANCE® Heparin (V. 02), RG_31_EN-U_4-10 Version 4.10, November 2020. 10. Siemens Healthcare Diagnostic Products, Germany. SYSMEX Automated Blood Coagulation Analyzer CS-2500 Reference Guide USA only, RG_36_EN-U_2-10, Version 2.10, September 2020. 11. Siemens Healthcare Diagnostic Products, Germany. Sysmex CS-2500 series Reference Guide, Application Sheet Heparin / INNOVANCE® Heparin (V. 05), RG_36_EN-U_2-11 Version 2.11, July 2021. Page 18 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 18 of 19  Bon Secours Mercy Health Richmond HealthPartners Laboratories Innovance Heparin Assay Sysmex CS-2500 and CA-660 COAG-0035 (Version 3.0) Approved and current. Effective starting 3/20/2024. Bon Secours HealthPartners Laboratories Richmond, Virginia Technical Section: Coagulation Procedure: Innovance Heparin Assay on Sysmex CS-2500 and CA-660 Appendix A. Best Practice for Double Centrifugation of plasma for coagulation samples sent to SMH core laboratory and/or to reference laboratory. Double centrifugation of the sample is to be performed to ensure that the sample is platelet poor. This is achieved by transferring the plasma, following the initial centrifugation process, to a properly labeled, non-activating sterile plastic centrifuge tube. ① The coagulation tech will receive 3 tubes from processing: a) An original sample tube b) A pre labeled pour-off tube c) A pre labeled freezer tube y p M o c :2 2 ② Use a plastic disposable pipette to transfer the plasma, from the centrifuged tube, to the pre labeled pour-off tube, then re-centrifuge the sample for an additional 10 minutes. A d e 24 11 may have collected at the bottom of the centrifuge tube. l l ③ When aliquoting to a secondary tube, take care not to include the residual platelets that 0 t r 12/1o 2/ 2 ④ Once re-centrifugation is complete, use a plastic disposable transfer pipette to transfer the platelets that have collected on the bottom). o on f double-spun plasma to the pre labeled freezer tube until ready to test (Do not include residual c ta n rren s If testing is not complete within the stated stability, the plasma must be removed from the cells and frozen. A frost-free freezer is not to be used. Frozen plasma samples must be rapidly U Cu thawed, at 37°C, gently mix the sample and perform testing immediately after thawing. If testing is delayed, after thawing, the sample may be held for two hours at 4°C until tested. ① ② ③ ④ *Note: buffy coat from each tube has not been disturbed. **NOTE: Heparin must be double centrifuged at a speed of approximately 2000 x g for 15 minutes.** Page 19 of 19 Uncontrolled copy printed by Leetta Bateman Lead MLS on 12/12/2024 11:22 AM (EST). Page 19 of 19