FAQ about suPARnostic® - suPARnostic® by ViroGates

suPARnostic® TurbiLatex The suPARnostic® TurbiLatex test has been validated on blood samples drawn in EDTA- or Lithium Heparin anti-coagulant collection tubes. Yes, only venous blood can be used. No, the suPARnostic® TurbiLatex test has not been validated on serum samples. It is not recommended to dilute samples as diluted samples report erroneous suPAR concentrations. If samples with a very high concentration must be estimated, please contact the manufacturer for guidance and support. As with all turbidimetric tests, a prozone effect can be induced, but with regards to the suPARnostic® TurbiLatex Test samples with a concentration up to 400 ng/ml do not interfere with the Test Measuring Range. The suPARnostic® TurbiLatex Reagents come in universal bottles and need to be transferred to original cassettes/bottles for the appropriate instrument before they are onboarded. No, dedicated cassettes/bottles for the clinical chemistry analysers of interest need to be purchased by the lab in addition to the TurbiLatex kit. The suPARnostic® TurbiLatex Reagents must be transferred to the dedicated cassettes/bottles before being placed in the appropriate instrument. The analysis of suPAR on the Chemical Clinical instruments takes 10 minutes. We do not have an FDA approval yet, but will apply for it. We are CE/IVD approved which is enough to be able to sell in Europe and many other markets. suPARnostic® Quick Triage Test For the suPARnostic® Quick Triage Test plasma in EDTA anti-coagulant collection tubes have been validated. The suPARnostic® Quick Triage test has not been validated on serum samples. Yes, only venous blood can be used. The following extra equipment is required; centrifuge, pipettes (10uL-100uL), tips, and tubes. The blood sample should preferably be centrifuged as soon as possible. If centrifugation is not possible directly after the blood is drawn then keep it on a stable surface and do not shake the tube as this could lead to mixing of haemolysed blood with the separated plasma. The blood sample is stable for approx. 12 hours (preferably kept at 2-8 degrees or at RT). If hemolysis occurs, the sample should not be used. Centrifuge the blood sample directly after collection at After the blood has been drawn plasma needs to be isolated by centrifuging at 3.000 g for 1 minute or until plasma and blood cells have separated. After isolation, EDTA plasma samples are stable for up to 24 hours at room temperature, three days at 2-8°C, and -20°C for longer periods. It is not recommended to dilute samples. To estimate higher concentrations, other tests should be used, such as the suPARnostic® AUTO Flex ELISA kit. Please contact the manufacturer for more information. Yes, as the reaction is not stopped, it is important to measure after 20 minutes, since the method and calibration curve are based on an incubation time of 20 minutes. Yes, when using the aLF reader for suPAR measurements all tests are considered as an individual test. About 1-2 min between each measurement should be enough to change the test device and scan the barcode. Make sure that every test device must be incubated for 20min. The method of choice is depending on your measuring rate. For example, if you only need to test one patient, the suPARnosticQT20 method is preferred as the Reader will measure automatically after 20 minutes. The reader cannot be interrupted during the incubation time, but the measurement will be done automatically. If you have several patients for testing, you can use the suPARnosticQT method. Here you need to measure the device manually after 20min of incubation time. this way more samples can be measured within one hour. No. The suPARnostic® Quick Test Reader is the older version. However, it is still possible to measure suPAR on this reader. The batch specific methods can be downloaded from the ViroGates support page. The aLF Reader is available for purchase through our webpage. To measure suPAR on the aLF reader, barcodes with the batch specific methods are included in the suPARnostic® Quick Test kit. The barcodes included in the suPARnostic® Quick Test kit is to be used together with the aLF Reader. If suPAR is measured on the suPARnostic® Quick Test Reader, the methods have to be downloaded from ViroGates support After the blood has been drawn, plasma needs to be isolated by centrifuging at 3.000 g for 1 minute. Then plasma is mixed with running buffer and added to the device, whereafter incubation for 20 minutes is needed. The reading of the test takes ~30 seconds. In total one, the test takes ~25 minutes. Multiple tests can be done simultaneously using the suPARnosticQT method. We do not have an FDA approval yet, but will apply for it. We are CE/IVD marked and which is enough to be able to sell in Europe and many other markets. suPARnostic® ELISA Yes, we provide more than enough of all components to run the entire plate once or separately 4 times using 3 strips of clear plate only. It is helpful when you have to measure less samples at once not using the whole clear plate. Due to the clinical impact of the results, we recommend testing in duplicates. However, if a lab has proven robustness between duplicates, the assay can then be run in singlets to provide 89 tests per plate. Robustness between duplicates is achieved through pipetting and washing accuracy, thus determined by the technical expertise of the testing lab – please refer to our detailed guide to running a suPARnostic® ELISA kit successfully. For up to 3-4 hours. The kit needs to be kept at room temperature for about 30 minutes before use. Yes, only venous blood can be used. It depends on the number of samples measured, but it normally takes about 2 - 2.5 hours. We do not have an FDA approval yet, but will apply for it. We are CE/IVD marked and which is enough to be able to sell in Europe and many other markets. Scientific Questions All individuals have a measurable suPAR level. In healthy blood donors (N9305) the suPAR level is around 2 – 3 ng/ml (25-75% interval from 1.76 – 3.23 ng/ml), in patients attending an ED around 3 – 6 ng/ml, and in patients with severe disease and organ failure, suPAR is often in the double-digits. The higher the suPAR level, the higher the risk of disease progression and the worse the prognosis. suPAR is not a diagnostic test for SARS-CoV-2. However, suPAR is a risk marker for whether a COVID-19 patient will develop respiratory failure or not. The higher the suPAR, the higher the risk of ending up intubated. Furthermore, suPAR has a high negative predictive value, meaning that patients with low suPAR, e.g. below 4 ng/ml, have very little or no risk of suddenly developing respiratory failure. Together with clinical scoring systems (e.g. Early Warning Score) and C-reactive protein levels, suPAR provides significant value to the decision of discharge or admission of COVID-19 patients. Inflammation is a vital part of the immune system's response to injury and infection. It is the body's way of signalling the immune system to heal and repair damaged tissue, as well as defend itself against foreign invaders, such as viruses and bacteria. Even though CRP and suPAR are positively correlated, suPAR has several advantages over CRP. suPAR is a stronger marker of readmission and mortality. Furthermore, acute medical patients with low CRP (<10 mg/L), can have elevated suPAR and these patients have high risk of readmission and mortality (Rasmussen et al, Emergency Medicine Journal, 2016). Observations in trials tell us it is within hours. A patient’s suPAR level continues to rise as the infection gets more serious. So, an upward trend is seen in slowly worsening conditions. However, although this is difficult to provide evidence for, it is likely that patients who have an elevated suPAR level before becoming infected, have a worse immune response to the infection and therefore progress more rapidly into severe sepsis. suPARnostic® can be safely used on pregnant women. There is a lack of data on infants. In children with pneumonia, a high suPAR level is associated with a lengthy stay at the hospital, suggesting that suPAR is also a marker of disease severity in children (Read the supporting this here) There is variation in suPAR levels between individuals and genetics may have some effect on the suPAR level. However, it is unlikely to have major influence on suPAR levels, as very few blood donors have suPAR levels as high as those who are severely ill (Read the supporting this here) Yes, any blood type can be used to measure suPAR. We continue to demonstrate the value of suPAR with studies and trials that only serve to provide repeatedly consistent results. Over 600 papers have been published on this matter. The suPAR level will decline after the infection or inflammation have been defeated. In individuals who stop smoking, suPAR levels taken after 4 weeks show a significant reduction (around 1 ng/ml) almost to the levels of non-smokers. How long this decline takes is unknown, but probably too slow to monitor whether a patient is responding to the therapy. It will likely take at least a week before a response in suPAR can be observed. suPAR is very stable in plasma and the sample can be frozen for up to a year. Avoid defrosting and freezing cycles The VG1 antibody binds to the suPAR DIII domain and the VG2 antibody binds to the suPAR DII domain. This means that the suPARnostic assays captures full length suPAR (D1D2D3) and truncated suPAR (D2D3). These are the two suPAR forms that carry strong prognostic value. General Questions suPAR (soluble uPAR) is a protein in the blood that reflects immune activation. All human beings have a baseline level of suPAR that is individually determined and increases with age. It is a universal and broadly applicable prognostic biomarker of chronic inflammation and immune activation across diseases. suPAR can be used for risk stratification of acute medical patients. Read more about suPAR here. suPAR stands for soluble urokinase plasminogen activator receptor. All individuals have a measurable suPAR level. In healthy blood donors (N9305) the suPAR level is around 2 – 3 ng/ml (25-75% interval from 1.76 – 3.23 ng/ml), in patients attending an ED around 3 – 6 ng/ml, and in patients with severe disease and organ failure, suPAR is often in the double-digits. The higher the suPAR level, the higher the risk of disease progression and the worse the prognosis. suPARnostic® is the only CE-IVD certified product range applied for clinical determination of suPAR (soluble uPAR) in human plasma and serum. Read more about the suPARnostic® products here. Biomarkers are a measure used to monitor and predict health states in individuals or across populations so that the appropriate therapeutic intervention can be planned. A wide range of biomarkers are used today. Many of these biomarkers are relatively easy to measure and are an important part of the routine medical examinations. Examples of biomarkers include both pulse and blood pressure measures as well as more complex laboratory tests of blood and tissues. The key issue at hand is determining the relationship between any given measurable biomarker and the relevant clinical endpoints. While a wide range of biomarkers are used today, suPAR in particular is a universal and stable marker that determines the health status of the patient. See a more elaborate comparison of suPAR and other biomarkers here. Plasma is the largest part of your blood, making up 55% of it. When separated from the rest of the blood, plasma is a clear light-yellow liquid. The main role of plasma is to take nutrients, hormones, and proteins to the parts of the body where it is need. The plasma also transports and removes waste products from the cells. No. Read more about the relevance of suPAR in other clinical areas here. Only a few general practitioners measure suPAR. Contact ViroGates for more info on this. No, not at the moment. We have direct sales in the Nordic countries, France, and Spain. We also sell the suPARnostic® products through distributors in a number of other countries. For more information or to purchase our products, please click here. Yes, it is possible. Adjusted analyses show that a 30-year-old non-smoking, and physically active male of normal weight on a healthy diet has a suPAR level of about 2.5 ng/mL, whereas a 30-year-old obese, heavily smoking, and inactive male on an unhealthy diet has a suPAR level of about 5.4 ng/mL. A change towards a healthier lifestyle is associated with a suPAR decrease. Hence, suPAR is not a death sentence – it is an early warning signal. Acute inflammation: Is a short-term response normally leading to recovery. Biomarker: A biomarker is a substance used as an indicator of normal biologic processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention. Chronic inflammation: Is a long-term, dysregulated response not leading to recovery, but rather, to tissue damage and increased disease severity. Inflammation: Is an immune response which clears infections caused by foreign pathogens, heals injured tissue, and restores homeostasis. If the inflammation does not resolve, it results in chronic inflammation. Membrane: The membrane of a cell is the interface between the cellular machinery inside the cell and the fluid outside. Protein: Biological molecule made out of chains of amino acids. Receptor: Molecule that binds another molecule (ligand). uPAR is receptor for uPA (see Ligand). ROC: Graphical plot of the sensitivity vs. (1 - specificity). Used in order to compare two or more different methods.