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1 Instructions for Use Suparnostic® Turbilatex Reagents 1 Instructions for Use suPARnostic® TurbiLatex Reagents INTERPRETATION OF suPAR suPAR level Clinical decision soluble urokinase Plasminogen Activator Receptor <3 ng/mL Supports the decision of discharge. T005 The underlying health condition is good, and the prognosis for surviving is very high. suPARnostic® and the ViroGates A/S ViroGates logo are Banevaenget 13 3-6 ng/mL Some disease activity or co-morbidity is present. registered trademarks of Birkeroed 3460, Some readmissions and an elevated mortality are ViroGates A/S Denmark. Denmark expected after 6 months follow-up. ©2008 ViroGates. Tel: +45 2113 1336 www.virogates.com All rights reserved. >6 ng/mL Attention is needed. High risk of mortality. Further tests are needed to find the presence of severe disease. Table 1: Simplified suPAR clinical decision scheme (data from Danish hospital)3. Interpretation of results must be made considering the patient´s clinical history and results of other diagnostic tests. The decision to discharge This product is protected by one or more US, European, and/or foreign can, therefore, not be performed solely based on the patients suPAR patents. value. The T005 product is validated on the automated Roche Cobas c701/2 PRINCIPLES OF ASSAY PROCEDURE ® instrument (Cobas is a trademark of Roche), and this instruction for use The suPARnostic® TurbiLatex test is a turbidimetric immunoassay is dedicated for the Roche cobas c701/c702 biochemistry analyser. that quantitatively determines suPAR in human plasma samples. The first stage of testing is an incubation of human origin specimen (EDTA- Refer to the webpage http://www.virogates.com for instructions for use or Heparin plasma) with the R1 reagent. After 5 minutes of incubation, for other biochemistry analysers and other languages. Alternatively, the R2 reagent is added, and the reaction starts. The reaction buffer R2 contact your local distributor for instructions in your language. is a suspension of latex particles coated with rat and mouse monoclonal antibodies to suPAR. After the R2 addition the process of suPAR INTENDED PURPOSE aggregation begins, the level of accumulation is determined by the For Professional Laboratory Use amount of scattered light during measurement of light absorption. The ® The suPARnostic TurbiLatex Reagents are used for automated linear calibration curve, created before the start of the test, is used to determination of soluble urokinase Plasminogen Activator Receptor determine the concentration of suPAR in human plasma samples. (suPAR) in human EDTA- or Heparin plasma in ng/mL on Roche cobas c701/2 analyzers. REAGENTS AND MATERIALS suPARnostic® is a prognostic biomarker of chronic inflammation and Reagents Provided; immune activation across diseases. suPARnostic® can be used for risk stratification of acute medical patients. • Reagent 1 – Dilution Buffer. • Reagent 2 – Latex Particle Reagent. suPAR IS A MARKER OF DISEASE PROGRESSION • Instruction for use. suPAR is the soluble form of urokinase Plasminogen Activator Receptor (uPAR). The amount of suPAR is a measure of immune activation and This kit contains ready-to-use R1 Dilution Buffer and R2 Solution of inflammation.1 suPAR is a biomarker which is increased by the presence Latex particles coated with anti-suPAR antibodies. The volume is and severity of disease. suPAR has a high negative predictive value for enough for a full cassette for the c701/2. ruling out disease progression. This means that patients with a low (<3 ng/mL) suPAR level have a good prognosis and a low risk of re- Product/Buffer R1 in mL R2 in mL No. tests admission and mortality3, supporting the decision of discharge of the T005 (c701/2) 75 28 420* patient. A high suPAR level (< 6 ng/ml) is a strong measure of chronic *no. of tests refer to the actual number of tests in the bottle, including dead volume inflammation and the underlying risk of negative outcomes, including but excluding 12 tests for calibration, when all reagent is transferred to the empty 2 short term mortality (in hospital, 30 days, or 90 days) supporting the cassette. decision of further examination of the patient. Material required but not provided The use of suPAR in clinical routine adds significant additional • suPARnostic® TurbiLatex Calibrators T002 knowledge to the standard assessment based on early warning score • suPARnostic® TurbiLatex Controls T003 systems and standard parameters in pre-admission of acute medical • Clinical chemistry analyser patients. Hence, suPAR is broadly applicable, e.g. in the Emergency • Adjustable pipette with tips, 20 200 Department, especially in relation to discharge decisions of patients, as μL – μL well as for identifying non-diagnosed inflammatory disease. • Disposable gloves A cluster-randomised interventional study showed that up- or down- triaging of patients based on suPAR levels increased the number of NUMBER OF DETERMINATIONS patients to be discharged (low risk) with 34%4 and reduced the number 1 mL of R2 reagent and 3 mL of R1 reagent is sufficient for 20 tests of hospital bed days5. when measured on Roche Cobas® c701/2 analyser. This does not include dead volume in the bottles. EXPECTED VALUES All individuals have a measurable suPAR level, and in healthy blood LOADING OF REAGENTS donors (N=9305) the median suPAR level for men age 18-65 years old The Reagents shall be transferred to the appropriate cassettes from is 2.22 ng/mL (25-75% interval from 1.76-2.90 ng/mL)6, for women 18- 65 years old is 2.56 ng/mL (25-75% interval from 2.05-3.23 ng/mL)6, Roche before they can be used on the analyser. patients attending Emergency Departments around 3-6 ng/mL2,3,7, and R1 reagent is transferred to position B and R2 into position C. For more in patients with severe disease and organ failure, suPAR is often in the information refer to the Roche Cobas c MULTI pack IFU. double-digits8, 9. The higher the level, the higher the risk of disease progression and the worse the prognosis. Note: suPARnostic® TurbiLatex Reagent 2 is on the Roche Cobas platform identified as R3. PI 175 Edition 1.0 June 2020 2 STORAGE AND HANDLING • Do not mix plasma samples from different patients or different Store at 2-8°C. blood samplings of the same patient. • Human samples may be contaminated with infectious agents. Do The suPARnostic Reagents is produced with 12 month shelf life from not ingest, expose to open wounds, or breathe aerosols. Wear protective gloves and dispose of biological samples properly. production date. Before use, check the expiry date on the label. • Be aware of possible dilution of suPAR in the case of a transfusion, The reagents have 8 weeks of onboard stability when kept at 2-8°C and infusion, or similar. monthly calibration performed. If not stored correctly the stability of the reagent may be affected and PERFORMANCE CHARACTERISTICS might not be efficient and misleading results may be obtained. Results presented below were obtained with the use of suPARnostic® TurbiLatex Reagents on the Roche Cobas® c702 analyser. The data SPECIMEN COLLECTION AND HANDLING shown is only valid for Roche Cobas c701/c702. Sample Type Sample Requirement Plasma sample 10 µL RESULTS Results are calculated by linear regression. If a different calculation SAMPLE COLLECTION AND STORAGE method than linear regression is used, the method must be validated. Collecting blood samples should be performed by trained and qualified Control the curve fitting by using suPARnostic® TurbiLatex Controls and staff in antiseptic conditions using approved venepunctures techniques. undertake corrective actions if results exceed the upper and lower limit. To prepare plasma samples, whole blood is drawn into a blood collection tube containing EDTA or Heparin anticoagulant. Centrifuge the blood at MEASURING RANGE 3,000 x g between 1 - 10 minutes until blood cells and plasma have The measuring range of the suPARnostic® TurbiLatex test is 1.8 ng/mL separated. to 16.0 ng/mL on a Cobas® c701/c702 analyser. It is not recommended to dilute samples above measuring range. Do not use hemolysed, contaminated, or hyper-lipemic sample specimens. ANALYTICAL LIMITS SAMPLE STABILITY LIMIT OF BLANK (LOB) It is preferred to analyse the samples as soon as possible, but EDTA- Three human plasma samples, diluted in 1:16 factor with water, were and Heparin- plasma samples are stable for: measured 20 times. In total, 60 blank measurements for 1 day. • 24 hours at room temperature (20-25°C). LIMIT OF DETECTION (LOD) • 3 days at 2-8°C. Five low concentration samples were tested in 12 replicates for 2 days. • -20°C for longer storage. Five freeze/thaw cycles over 5 days do not have an impact on suPAR The LOD for suPAR was determined in accordance with the guidelines concentration in a sample. of CLSI (Clinical and Laboratory Standard Institute)10 protocol and with proportions of false positives (α) less than 5% and false negatives (β) CALIBRATION less than 5%; based on 120 determinations, with 60 blank and 60 low- Together with suPARnostic® TurbiLatex Reagents, the suPARnostic® level samples. TurbiLatex Calibrators T002 should be used for calibration. The laboratory shall work out their frequency of calibration, but it is LIMIT OF QUANTIFICATION (LOQ), LOQ=LOD recommended to repeat the calibration at least once a month, and it is Acceptable Total Error = 30% to suPARnostic® ELISA. required to recalibrate when a new batch of TurbiLatex Reagents are LOB LOD LOQ used. Result 1.0 ng/mL 1.3 ng/mL 1.3 ng/mL Limit Result <LOB LOB < Results ≤ LOD Result ≥ LOQ QUALITY CONTROL Report Analyte not Analyte present, cannot Analyte can be Quality control of suPARnostic® TurbiLatex Reagents should be detected; be quantified; estimated ≥ 1.3 ® performed with suPARnostic TurbiLatex Controls T003 after each Concentration < concentration < 1.3 ng/mL ng/mL calibration. Quality control ranges and limits should be fitted to the 1 ng/mL specific laboratory’s experience and knowledge.
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