EXPECTED VALUES All individuals have a measurable suPAR level, and in ACCURACY (METHOD COMPARISON) PRINCIPLES OF ASSAY PROCEDURE healthy blood donors (N=9305) the median suPAR level Passing-Bablok correlation toward suPARnostic® ELISA The suPARnostic® Quick Triage is a lateral flow for men aged 18–65 years old is 2.22 ng/mL (25-75% has been performed to estimate the Quick Triage ability immunoassay. The device utilizes monoclonal rat and interval from 1.76–2.90 ng/mL)2, and for women 18-65 to quantify suPAR in patient samples. Instructions For Use gold-conjugated mouse antibodies against human suPAR years old 2.56 ng/mL (25-75% interval from 2.05–3.23 Results suPARnostic® Quick Triage for aLF Reader to give a quantitative measurement of the plasma suPAR ng/mL)2. In patients attending emergency departments Sample No. of Slope Y-intercept Passing- Range level. The EDTA- or heparin-plasma is mixed with running type Pair Bablok Value soluble urokinase Plasminogen Activator Receptor 3,4,7 ® the suPAR level is around 3-6 ng/ml and in patients correlation Test Device buffer and applied to the suPARnostic Quick Triage with severe disease and organ failure, suPAR is often in Plasma 60 1.13 -0.39 0.893 1.3-18.7 device. During the 20 min. of incubation, the plasma A003 the double-digits5,6. The higher the suPAR level, the X = suPARnostic ELISA Y = suPARnostic Quick Triage sample reacts with gold-conjugated anti-suPAR higher the risk of disease progression and the worse the antibodies and migrates through the nitrocellulose WASTE HANDLING suPARnostic and the ViroGates A/S prognosis. Banevaenget 13 membrane. Gold-conjugate containing sample suPAR is Discard unused reagents and waste in accordance with ViroGates logo are registered trademarks of Birkeroed 3460, bound by a capture suPAR antibody at the Test line, country, federal, state, and local regulations. PERFORMANCE CHARACTERISTICS ViroGates A/S Denmark. Denmark while non-suPAR bound antibody is captured by the Limit of Blank (LOB) - show the variation of a blank ©2008 ViroGates. Tel: +45 2113 1336 Control line (anti-mouse antibody). sample (buffer only). Highest value from 3 batch Skype: Virogates The suPARnostic® Quick Triage is calibrated against an validations. Limit of Detection (LOD) - is the lowest internal control. No international standard have been possible detection of suPAR that is not a blank sample. established. Highest value from 3 batch validations. Catalogue LOT No. Consult Temperature no. (Batch No.) instructions for use Limits Limit of Quantification (LOQ) - is set to be the sample REAGENTS AND MATERIALS This product is protected by one or more US, with the lowest concentration in the range 0-2 ng/ml to European, and/or foreign patents. Reagents Provided have a CV% that does not exceed 25%. Highest value This kit contains reagents sufficient to perform 25 tests from 3 batch validations. devices. Do not re- Do not use Contains sufficient Use by Refer to www.virogates.com for further instructions and LOB LOD LOQ 1. Lateral Flow Devices, each in aluminum pouch use if package for
2. Assay Running Buffer, PBS buffer, pH 7.2, with Imprecision & Repeatability INTENTED USE REFERENCES proprietary additives and 0.05% Bronidox® as Intra-serial results are estimated on 5 measurements in 1. www.alf-reader.com For professional use. preservative. one day and provide a Mean, Standard Deviation, and 2. Haastrup E, Grau K, Eugen-Olsen J, Thorball C, Kessing LV, Ullum H: The suPARnostic® Quick Triage test is used for Soluble urokinase plasminogen activator re-ceptor as a marker for Quantity: 3.5 mL. Preparation: Ready to use. CV%. The inter-serial variation is between 5 days. The determination of soluble urokinase plasminogen use of antidepressants. PLoS One 2014, e110555. 3. Instruction for use. highest CV% from 3 batches are displayed below. 3. Raggam RB, Wagner J, Pruller F, Grisold A, Leitner E, Zollner- activator receptor (suPAR) in human EDTA- and heparin- 4. Barcodes to upload the methods. Schwetz I, et al: Soluble urokinase plasminogen activator receptor plasma in ng/ml. The suPARnostic® Quick Triage test is SAMPLE SAMPLE SAMPLE SAMPLE predicts mortality in patients with sys-temic inflammatory 1 response syndrome. J Intern Med 2014, 12238.:10. measured on the aLF Reader from Qiagen. 1 2 3 4 Material required but not provided X (NG/ML)= 2.0 4.0 7.4 14.0 4. Haupt TH, Petersen J, Ellekilde G, Klausen HH, Thorball CW, Eugen- WITHIN RUN 22% 23% 12% 10% Olsen J, et al: Plasma suPAR levels are associat-ed with mortality, Interpretation of results must be made considering the • Adjustable pipette with tips, 10 μL – 100 μL. CV (%) admission time, and Charlson Comorbid-ity Index in the acutely • Clock, Timer, or stopwatch. admitted medical patient: a pro-spective observational study. Crit patient´s clinical history and results of other diagnostic BETWEEN RUN 29% 20% 18% 18% CV (%) Care 2012, 16:R130. tests if available. • Disposable gloves. 5. Koch A, Zimmermann HW, Gassler N, Jochum C, Weiskir-chen R, • aLF Reader (#ESLR12-MB-6401). Bruensing J, et al.: Clinical relevance and cellular source of elevated LINEARITY soluble urokinase plasminogen activator receptor (suPAR) in acute SUMMARY OF suPAR AS A MARKER OF DISEASE • Eppendorf tube or other mixing tube. The Quick Triage analysis of suPAR on the aLF Reader, liver failure. Liver Int 2014, 10 PROGNOSIS has been demonstrated to be linear from 2.5 ng/ml to 6. Donadello K, Scolletta S, Taccone FS, Covajes C, Santono-cito C, suPAR is the soluble form of urokinase plasminogen REQUIRED TRAINING Cortes DO, et al: Soluble urokinase-type plasmino-gen activator 15.2 ng/ml, within 7.5% degree of nonlinearity in this receptor as a prognostic biomarker in critical-ly ill patients. J Crit activator receptor (uPAR). The amount of suPAR is a To use the suPARnostic® Quick Triage it is required that interval. Care 2014, 29:144-149. measure of immune activation and inflammation. suPAR the user is fully trained in how to operate the aLF Reader. 7. Rasmussen LJH, Ladelund S, Haupt TH, Ellekilde GE, Eugen-Olsen J, is a non-specific biomarker which is increased by Andersen O. Combining National Early Warning Score With Soluble HOOK EFFECT Urokinase Plasminogen Activator Receptor (suPAR) Improves Risk presence of disease. The higher suPAR level, the higher The suPARnostic® Quick Triage shows no prozone effect Prediction in Acute Medical Patients: A Registry-Based Cohort the risk of disease progression and the worse the in concentrations below 70 ng/mL (this is the highest Study. Crit Care Med. 2018 Dec;46(12):1961-1968. patient's prognosis. tested suPAR concentration).
Page 4 PI 303 v.1.3 April 2020 Page 1 PI 303 v.1.3 April 2020 REAGENT PRECAUTIONS AND RECOMMENDATIONS DEVICE DESCRIPTION RUNNING A TEST The aLF Reader it will automatically display if any error • For professional use. 1. To start a new test, touch the “New Test” field on has occurred during measurements. The reader has an • Do not use kit components beyond the indicated kit the touch screen display. internal quality control which is run every time the expiration date. 2. Scan either the suPARnosticQT or the reader is turned on. • Do not mix reagents from different kit lots. suPARnosticQT20 barcode, using the internal 2D- • Do not freeze any of the kit components. barcode scanner on the aLF Reader, provide in the • Do not mouth pipette or ingest any of the reagents. kit depending on the preferred measuring method. CALCULATION OF RESULTS ® • Do not smoke, eat, or drink when performing the NOTE: keep the barcode in a vertical angle. The suPARnostic Quick Triage device has to be used with
assay or in areas where samples or reagents are 3. Test method name (1), Lot ID (3), and Sample the aLF reader to give correct values. The user cannot
handled. Settings (5-7) will automatically appear on the evaluate the results by visually inspecting the Quick ASSAY PROCEDURE • Do not mix plasma samples from different patients screen. Triage device. The aLF Reader automatedly performs the It is critical that the volumes pipetted and the incubation or from different blood samplings of the same 4. Read the 2D-barcode with the Patient ID or write calculation of suPAR levels. time is followed precisely as described in this procedure. patient. the Patient ID manually. The aLF reader scan the test- and control line and Two measuring methods are offered for each batch. The determine the intensity of the lines. The calculation to • Human samples may be contaminated with suPARnosticQT method starts measuring the suPAR level infectious agents. Do not ingest, expose to open estimate the suPAR value is based on the test line. The when the ’forward’ button is pressed. wounds, or breathe aerosols. Wear protective aLF Reader uses a batch specific method for each batch The suPARnosticQT20 method measure the suPAR level of Quick Triage test devices for the calculation. The batch gloves and dispose of biological samples properly. 20 min after the button have been pressed. ’forward’ specific method is included in the test device kit as a QR • Do not use a device if the pouch is damaged or This allows the user to insert the device in the reader opened in some way. code. The method contains a calibration curve the reader during the incubation and ensures the incubation time is uses to convert the T- • Be aware of possible dilution of suPAR in the case Line’s intensity to ng/ml suPAR. correct. The mathematical calculation is made with a linear curve of transfusion, infusion, or similar. based on 6 reference samples with known
suPARnosticQT suPARnosticQT20 concentrations and a buffer only sample. REAGENT STORAGE AND HANDLING 1. Transfer 100 µl of running buffer to an empty tube. Devices must be stored in the sealed foil pouches. 2. Transfer 10 µl of plasma sample to the tube Store kit components at 18 24°C. LIMITATIONS OF TEST – containing 100 µl of running buffer. 5. Open tray on right side of the reader. Insert the The Devices and running buffer may be used until the Clinical diagnosis should not be based on the result of the Vortex the mixture or use the pipet to mix up and device with patient ID to the left and the sample date printed on the pouch or bottle. suPARnostic® Quick Triage test alone. Interpretation of down. well to the right after adding the patient sample. Tightly close the cap after each use. 6. results must be made considering the patient’s clinical 3. Transfer 60 l of the diluted sample to the well of Touch the “forward” button to proceed and IMPORTANT: Devices have to be used right after opening μ history and results of other diagnostic tests, if available. the suPARnostic® Quick Triage device. confirm that the cassette has been inserted in the the pouch. Cannot be stored for later use. correct orientation. 4. Incubate the device for 4. Scan the 7. The suPAR result will be displayed in ng/ml. The substances listed below were tested for interference 20 min on the table and suPARnosticQT20 ® SPECIMEN COLLECTION with the suPARnostic Quick Triage test. None of the insert the device into the method barcode. Insert 8. The suPAR value should be within the range 2-15 Sample Type Sample Requirement tested substances interfered with the performance of aLF Reader before the device in the aLF ng/ml. If the result is out of range, it will be shown Plasma Sample 10 µl of Plasma the test. uploading the method. (If Reader for incubation as < 2.0 ng/ml or >15 ng/ml, and the value cannot the user is NOT present and touch forward to SAMPLE COLLECTION AND STORAGE “ ” be considered as accurate and precise. If the Substance Concentration mmol/ L during the incubation it is activate the 20-minute Preparation of plasma samples; display shows INVALID, an error has occurred Bilirubin 0.10 – 0.50 recommended to use the incubation. 1. Whole blood is drawn into a centrifuge tube Hemoglobin 0.00 0.94 suPARnosticQT20). during the measurement. Re-run the sample and if – containing EDTA or heparin anti-coagulant. the result is INVALID again, check the extensive Triglycerides 0.00 - 23 5. Press “forward” to read 5. aLF Reader reads the 2. Centrifuge the blood at 3,000 x g for 1-10 min. 3. Transfer and store plasma samples in separate the device with the aLF device automatically instruction on the internet, or contact ViroGates Rheumatoid factors: marked tubes. Reader. Use the designated after 20 min. for support on phone number +45 2113 1336 or by 4. Date and identify each sample. For long-term batch method. email to [email protected] Samples from 16 patients with increased rheumatoid
storage, keep at -20oC. Avoid freeze/thaw cycles. factor in the concentrations (0-1600 kIU/L) were QUALITY CONTROL analyzed. No significant correlation to rheumatic factors Grossly hemolyzed, lipemic, or microbiologically The suPARnostic® Quick Triage uses the C-Line as the (R2=0.33) were observed. contaminated samples should not be used. Samples with internal quality control. The result is faulty if the C-line abnormally elevated levels of hemoglobin or bilirubin does not appear on the device after an otherwise may interfere with assay performance and sensitivity. successful run of plasma sample.
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