- - inhibition e.g. by aspirin - deficiency invWF (vWD) - GpIb deficiency (Bernard-Soulier syndrome) RISTOtest: Different disorders can lead to low aggregation in aggregation tendency ofvWF (vWS type IIb). aggregation in RISTOtest low mayindicate an enhanced strong aggregation response. A higher than expected (0.2 mg/ml) is applied which normally does not induce a In RISTOtest low alower concentration ofristocetin receptorsvWF. or RISTOtest high can be based on a deficiency ofGpIb in aggregation Attenuated aggregation. platelet (0.77 mg/ml) is applied, which normally induces a strong In RISTOtest high ahigh concentration of ristocetin in the individual patient sample. whether additional vWF improves or corrects RISTOtest the addition of vWF reagent in order to determine In the RISTOconfirm RISTOtest is performed with RISTOconfirm assay. RISTOtest can be applied in two concentrations activationaggregation. and to the platelet GpIb receptor and triggers platelet complex withvWF fromthe sample. This complex binds a forms Ristocetin ristocetin. contains reagent RISTOtest Background aggregation studies on the Multiplate For in vitro research use only. Reagent for use in platelet Intended use reagent. This box insert is valid for kit format MP0548 of vWF useonly For invitroresearch

vWF reagent for platelet aggregation studies aggregation for platelet Munich - Germany DiagnosticaVerum GmbH Valid for REFMP0548 vWF reagent kit reagent vWF

® analyzer

1 in

V.1.0-US-RUO Revised 2012-02 to low aggregation inRISTOtest. This may facilitate the differentiation of disorders leading unchanged. remained disorders other the whereas corrected, By theaddition of vWF reagent a deficiency ofvWF is of all waste materials according to the local regulations. exposure of specimen and reagents tothe skin. Dispose specimen and all materials, e.g. wear gloves, minimize General precautions should be followed when handling Warnings and precautions position. upright an in stored be should not transferred into micro test tubes, the original vial reconstituted reagent at 2-8°C. Ifreconstituted reagent is vial labelwhen stored under these conditions. Store the The reagent is stable until the expiry dateprinted on the Unopened vials of vWF reagent must be stored at 2-8°C. Storage and stability daily use. at least 100 µl of the reagent into micro test tubes for To achieve maximum stability after reconstitution, pipette temperatures. advisable to minimize exposure to light, air and elevated Keep all vials tightly closed when not in use. It is the vials was replaced by aninert gas. Note: solution shouldclear beand colourless. minutes and swirl gently tomix –donot shake!The 7.5 U/ml). Allowto stand atroom temperature for 10 (distilled or deionized) water (vWF concentration: Reconstitute the vWF reagent with 1.0 ml of high purity preparation Reagent aliquotation. lyophilised (7.5 U/ml) with 5 micro test tubes for The reagent is provided in the following format: Reagents * theoretical, not proven yet REF MP0548 –vWF reagent: REF vWF reagent; 1x 1.0 ml, RSOetRISTOconfirm RISTOtest vWD vWD Thrombocytopenia Aspirin syndrome) (Bernard-Soulier GpIb deficiency Normal Due torisk minimization procedures the vacuum in abnormal normal abnormal abnormal abnormal abnormal abnormal abnormal * normal normal

Test procedure for RISTOtest with addition ofvWF and the box insert ofRISTOtest. analysis. before blood the preheat not Do samples. refrigerate complete mixing of the content. Do not freeze or collection. Gently invert the sample tube to ensure Avoid foam formation inthe sample tube during blood hirudin) isemployed. or citrate heparin, (i.e. anticoagulant sample same an individual sample with reference collectives when the each centre. The blood collection system must bestandardised at order to avoid enhanced citrate levels. in volumes, fill indicated the to filled are tubes collection citrate) may be used. Always ensure that citrated blood (3.2% tubes citrated or tubes lithium-heparin standard Alternatively commercial hirudin tubes (MP0600), ofblood). 3ml for solution hirudin µl 30 (e.g. applied into the blood collection tube in a ratio of 1:100 hirudin is diluted toa concentration of 2.5 mg/ml and final concentration of 25 µg/ml. For this recombinant hirudin assample anticoagulant is recommended with a significantly affects results of the tests The anticoagulant used for the blood collection Sample collection additionvWF). of with these obtained in RISTOtest high (without the compared be should RISTOconfirm in obtained Results Quality control such as methyl ester. Furthermore the saline must not contain any additives skew results. saline diluent solution or shorter incubation times may of non-preheated The use procedure. this exactly Follow Final concentration of ristocetin: 0.77 mg/ml Final concentration of vWF: 0.23 U/ml (RISTOconfirm) reagent in the Multiplate instructions the Follow collection. blood after hours 0.5-3 of period the within analyzed be should Samples ofthe analysis Performance  + 50 µlRISTOtest reagent  room temperature) anticoagulated blood orcitratedblood, + 300 µlwhole blood (hirudin- / heparin- + 280 µl saline 0.9%, preheated at 37°C 20 Start test 3minutes incubation

µl It is only possible to compare the results of vWF reagent (7.5 U/ml) ® user manual, short instructions manual manual instructions short manual, user  6 minutes measuring time time measuring 6 minutes

2 . The use of [email protected] www.diapharma.com 800-447-3846 Service Technical Customer Service 800-526-5224 OH 45069, USA Chester West Road, Beckett 8948 DiaPharma Group, Inc. Distributor [email protected] www.multiplate.us Phone: +49-89-125556-0 MunichGermany - Verum Diagnostica GmbH Manufacturer 781-8. 96(6): 2006; Haemost measure platelet aggregation in whole blood. Thromb Multiple electrode aggregometry: A new device to 2 ThrombHaemost 2008; 99(1): 121-6. aggregometry before and after clopidogrel treatment. aggregometry andmultiple electrode platelet ADP-inducedplatelet aggregation withlight transmission of Assessment N. Beckerath von A, Kastrati A, Schömig 1 Literature Sibbing D, Braun S, Jawansky S, Vogt W, Mehilli J, J, Mehilli W, Vogt S, Jawansky S, Braun D, Sibbing Tóth O, Calatzis A, Penz S, Losonczy H, Siess W.