Application Sheet for Rivaroxaban (Xarelto ) Standard Range with. BIOPHEN Heparin LRT (#221011/221013) RUO

Transcription

1 Instrument Adaptation The attached instrument adaptation has been prepared and validated by the reagent manufacturer, Hyphen-Biomed. Instrument Siemens BCS-XP Product BIOPHEN Heparin LRT Analyte Rivaroxaban Range Standard Range SKU# , , , Aniara SKU# A , A , A , A Document# D750-07/BI/ (Riva SR)-BCS-XP v1 Please note that the reagent/instrument adaptation described in the following pages has not been approved or cleared by the US FDA. In the US, it is intended For Research Use Only. Not for Use in Diagnostic Procedures. Aniara Diagnostica LLC 7768 Service Center Drive West Chester, OH Phone (513) Fax (513) com

2 Page 1 of 10 Application Sheet for Rivaroxaban (Xarelto ) Standard Range with BIOPHEN Heparin LRT (#221011/221013) This application sheet defines the measurement procedure and settings for Rivaroxaban Standard Range using kit BIOPHEN Heparin LRT to optimize product performance on Siemens BCS-XP. Modification of parameters on your system may affect performance and results. If the application is changed, it is responsibility of the user to validate any modifications and their impact on all assay results. For additional information, please refer to the IFU of analyzer and reagents. A calibration curve and a specific configuration are required. Test Principle BIOPHEN Heparin LRT assay is a kinetic method based on the inhibition by Rivaroxaban of Factor Xa (FXa), which is at a constant concentration and in excess. The remaining FXa is then measured by its amidolytic activity on a Factor Xa specific chromogenic substrate, which releases pna. The amount of pna generated is inversely proportional to the concentration of Rivaroxaban in the tested plasma. Rivaroxaban sample + Factor Xa excess > [FXa - Rivaroxaban] + Factor Xa residual Factor Xa residual + Substrate Factor Xa > Peptide + pna Assay synopsis 40µL sample diluted in NaCl Dilution Factor 1/ µl Hep L R1 Intensive sec at 37 C 100 µl Hep L R2 Special/SCS cleaner Measure sec at 37 C OD/min at 405 nm Calibration Lin-Log (concentration-od/min) 1. Preparation of BIOPHEN Heparin (LRT) reagent (Ref ) No preparation is required, all the reagents are liquid, ready to use Reagents provided: R1 : SXa-11 Substrate (suggested ID on analyzer: Hep_L R1) R2: Factor Xa (suggested ID on analyzer: Hep_L R2) Let stabilize 30 min at RT (18-25 C) while shaking the vial from time to time and then 30 min on board of the analyser before use. Homogenise the content before each use. Reagents are stable 6 months at 2-8 C, 14 days at RT (18-25 C). Reagent not provided: Diluent: Saline NaCl 0.9% (additives free). (suggested ID on analyzer: NaCl) Distilled water (additives free)

3 BIOPHEN Heparin LRT(#221011/221013)- Rivaroxaban Standard Range Siemens BCS-XP System Application Page 2 of 10 (LM 2. Preparation of controls and calibrators Reagent available at Hyphen BioMed: BIOPHEN Rivaroxaban Plasma Calibrator (Ref: ) (suggested ID on analyser: Riva Cal1...) 3 levels of ready to use calibrators. BIOPHEN Rivaroxaban Control Plasma (Ref: ) (suggested ID on analyser: Riva CQ1 ) 2 levels of ready to use controls. Reconstitute each calibrator level and control with exactly 1mL of distilled water. Homogenise the content before each use. Let stabilize 30 min at RT (18-25 C) while shaking the vial from time to time and then 30 min on board of the analyser before use. Controls and calibrators are stable 7 days at 2-8 C or 48 hours at RT (18-25 C), and 2 months at -20 C. 3. Stabilization, Storage and Use : Before use: Let the reagents stabilize for 30 minutes at room temperature (18-25 C), while shaking from time to time. For homogenous reactivity of the assay, it is necessary to let the reagent s temperature stabilize for at least 30 minutes on-board of the analyser before any use. It is recommended to run the calibration curve with a freshly reconstituted calibrator. Homogenise well the reagents, calibrators and controls before each use. Storage: Ensure vials are closed with their original specific caps before storage at 2-8 C. Cautions : data indicated in the reagent s pack insert are validated with vials kept closed, avoiding any evaporation or contamination. must be verified and adjusted, according to the laboratory exact working conditions. Any reagent of biological origin must be handled with all the required cautions, as being potentially infectious. Do not interchange the reagents from different lots. Quality controls must be run regularly, and for each new batch of reagents, after an important maintenance of the instrument, or if measured values are not in compliance with the one expected for the method. Performances may present slight variations according to the instrument used. Each laboratory can determine its own acceptation range in relation to the working conditions. The calibration curve is validated when linearity, as well as measured control values, are in compliance. Assay Calibration must be repeated after reagent lot change, important analyser s maintenance or when controls values fall out of the acceptance range (after checking all system parameters).

4 BIOPHEN Heparin LRT(#221011/221013)- Rivaroxaban Standard Range Siemens BCS-XP System Application Page 3 of 10 (LM 4. Results: The calibration curve type is: Log (OD/min) - Lin (concentration). The sample s Rivaroxaban concentration is directly calculated from the calibration curve The results are expressed as ng/ml of Rivaroxaban The sample of high concentration can be diluted in a normal plasma pool. Low concentration samples can be tested using the low range calibration. Duplicate testing is recommended. 5. Performances Characteristics: Precision Within-run Mean value ng/ml CV (%) Rivaroxaban QC Rivaroxaban QC Within run precision was calculated with Rivaroxaban C1 and C2 in 10-fold determination. Measuring Range Chromogenic Assay Measurement Principles Rivaroxaban (ng/ml) BIOPHEN Hep LRT Riva (OD/min) Analytical measuring range Chromogenic Method calibrated from standard curve using BIOPHEN Rivaroxaban Calibrator Chromogenic Method raw values ng/ml** ** ** These values are given for information only and may vary from lot to lot and instrument to instrument. For low samples the assay with low range configuration is recommended. Method comparison (ng/ml): Method Comparison r slope Interference Studies No interferences up to... Intralipids / Chyle [mg/dl] Haemoglobin [mg/dl] 1200/1000 Bilirubin (F) [mg/dl] Bilirubin (C) [mg/dl] 60

5 Page 4 of 10 Calibration Curve (Example use only) BIOPHEN Calibrator Plasma (Rivaroxaban ng/ml) OD/min A new calibration curve should be established with each change of reagent lot or with any deviation from controls or proficiency testing limits. The curve below is given as example only.

6 1. Assay parameter set up Creation of reagents. From the computer desk, open «Data definition tool». In Menu Definitions / Reagents, check if the products do not already exist If necessary, click on icon «new» and create the 2 reagents and the diluent following the instructions below. Page 5 of 10 Name : HEP LRT SUBSTRAT R1 Abbreviation : SUBHEPLRT Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring Name : HEP LRT FXa R2 Abbreviation : XAHEPLRT Wash when reagent is changed special Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring Name : NaCl Abbreviation : NaCl Wash when reagent is changed low Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring To be filled by the user according to working conditions, always use the same working conditions (refrigerate or not). Enter the 4 digits of the code assigned by the user to the product. Then click on Add Place reagents in original vials in the following types of racks: Kit reference Reagent Rack SUBHEPLRT* 50### XAHEPLRT* 50### NaCl NaCl 10### *use a reducer Note: always use the same type of racks (refrigerate or not) as performances may vary.

7 Standard / Calibrators and controls set up: Create the calibrator plasma according to the here below information: In the Menu: Definition/ standards calibrators It is necessary to create each level of the calibrator set separately. Name : Rivaroxaban Cal1 Abbreviation : Riva Cal1 Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring Name : Rivaroxaban Cal2 Abbreviation : Riva Cal2 Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring Page 6 of 10 Name : Rivaroxaban Cal3 Abbreviation : Riva Cal3 Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring To be filled by the user according to working conditions Enter the 4 digits of the code assigned by the user to the product. Then click on Add Create the controls according to the here below information: In the Menu: Definition/ Controls It is necessary to create each level of the control set separately.

8 Page 7 of 10 Name : Rivaroxaban Control1 Abbreviation : Riva Ctl1 Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring Name : Rivaroxaban Control2 Abbreviation : Riva Ctl2 Enzyme blank value measurement with bottle change Duration : 0 sec. Continuous stirring Assay Procedure definition In Data definition tool, Menu Definitions / Assay procedure enter the following parameters: General Identification Date of last change * Name RivaHLRT Number 2047 Measure enzyme blank value every 0 hours Finish measurement after 60 Sec or 0 ma Measurement speed Mixing intensity Wavelength Primary absorbance range n-replicate(s) Normal Normal 405 nm ma Permitted CV Samples and controls 1 [4] 20 % Calibration / EBV 2 [4] 10 % Raw values merging Arithmetic Mean Dilution Factor 1 : 10 [5] (for all measurements of this assay procedure) Dilution medium NaCl

9 Page 8 of 10 Evaluation and checking methods: AUC2 E Several evaluation methods and checking methods may be selected. (Double click = select/deselect) cmax E Delta A E Delta A per minute E Legend: refer to operator manual for symbols meanings Drifting baseline E [E] = Evaluation method Fixed absorption E [V] = Checking method Parameters of method: Delta A per minute Start time 20 sec Stop time 50 sec Correlation coefficient 0.95 [3] to be filled and adjusted by the user depending on the analyser [4] to be filled and adjusted by the user. It is recommended to perform the tests in duplicate. (1=simple et 2= double) [5] no dilution =1 Click on «Pipetting» and enter the following pipetting sequence: Click on Add a new cycle : a row will appear in the upper part of the table. Add the row for each action in the lower part of the table and complete all rows related to each cycle before adding a new cycle. Pipetting cycle Add new cycle Delete last cycle Cycle Transfer Arm Final washing Rotor / Transfer action Sample Arm Reagent Arm Reagent Arm normal normal normal Prioritize undiluted Incubation No mixing / Start measurement Time window [sec] min. max. Pipetting Media Add new row Delete last row Cycle Pipetting medium / target Reagent Speed Volume (µl) N Air Sample Outer cuvette Air Reagent Outer cuvette Air Reagent Inner cuvette SUBHEPLRT XAHEPLRT Assay Definition: In Data definition tool, menu «Definitions / Assays» click on new and enter the following parameters: General identification Date of last change Assay Name RivaHLRT Assay Number 2045 Host ID * Host result ID * Version *

10 Screen display Shorts codes in lists RivaHLRT Assay priority 1 Internal result unit ng/ml User-defined result unit and number format ng/ml One internal units is 1.0 User-defined units Host unit ng/ml One internal unit is 1.0 Host unit Result Factor 1.0 Selection of the procedures of tests of valuation methods and check RivaHLRT E Delta A / minute + - Formula to calculate the raw value : Minimum Arithmetic Mean Unit of raw value : ma/min Page 9 of 10 Results determination Reference range / Cut-off 0 to 0 ng/ml without reference curve Evaluation method Calculate result based on ref. curve of another assay Assay selection calculate result based on own reference curve Calibration procedure regression x lin y log Minimum raw value 0.0 Extrapolation limits : (0,01-1,0)* lowest concentration 1.8 [6] (1,0-2,0)* highest concentration 1.0 [6] Recording curve Calibrate using master curve Calculate using measured reference curve Tube curve validation activated Permitted deviation from the tube curve % No. of automatic repeats of invalid reference curve points 0 Calibration with calibrator set Calibration with automatic dilution of a standard Calibrator Desired Concentrations : Riva Cal1 * Riva Cal2 * Riva Cal3 * Add calibrator Remove calibrator Control assignment [6] this may need readjustment depending on the expected extrapolation (<1.0 when measuring activity < lowest point of the curve and >1.0 when measuring activity > highest point). [7] to be filled and adjusted by the user according to the lot used. Click on control assignment in order to set the control: type, acceptance range, automatic frequency Configuration and lot specific information Configuration: Close «Data definition tool», and open BCS XP SW Menu Definitions / configuration: in the left column, select Assays

11 Page 10 of 10 In the left list «Assays», select «RivaHLRT» then click on icon to transfer the assay in the right column «Selection» to activate the assay. Option: it is possible to organise the Assay display by moving the assay s name in the list Selection : use the icons. Lot specific information: In Menu Definitions / Lots info: enter the product s codes and lot numbers.