Cold storage of citrated whole blood induces drastic time-dependent losses in factor VIII and von Willebrand factor: potential for misdiagnosis of haemophilia and von Willebrand disease

Abstract

This study investigates the effect of pre-analytic storage conditions on the laboratory evaluation of von Willebrand disease (VWD) and haemophilia. Samples from healthy controls and patients with VWD were stored as whole blood and as separated plasma, both at room temperature and on crushed ice, for two different time periods (3 or 6 h). In samples from healthy individuals (n=10) and in patients with suspected type 1 VWD (n=10), storage of whole blood on ice caused a drastic time-dependent decrease in von Willebrand factor (VWF):ristocetin cofactor activity, in VWF:antigen activity and factor VIII activity (mean+/-SD) to 35+/-18, 55+/-23 and 53+/-15% of baseline levels after 6 h storage, respectively. Patients with type 2 VWD and non-detectable VWF:ristocetin cofactor activity did not demonstrate such drastic cold-induced losses in VWF and factor VIII levels. Storage of plasma caused only minor changes in VWF levels. The cold-induced loss in VWF might thus depend on the presence of platelets and of high molecular weight VWF. Chilling of platelets induces a clustering of the glycoprotein Ib subunit. We therefore hypothesize that cold-induced loss in VWF might be due to a cold-promoted binding of VWF to glycoprotein VWF receptor Ib alpha. These results suggest a serious potential for misdiagnosis of haemophilia or VWD due to inappropriate pre-analytical handling of blood.

Influence of 8 and 24-h storage of whole blood at ambient temperature on prothrombin time, activated partial thromboplastin time, fibrinogen, thrombin time, antithrombin and D-dimer

“…The storage-induced increase of APTT is most likely due to the instability of coagulation factor VIII as shown by other studies [5,9]. APTT reagents exhibit different sensitivities for coagulation factor deficiencies, which might be the cause for the different study findings.…”

The diagnosis and management of von Willebrand disease: a United Kingdom Haemophilia Centre Doctors Organization guideline approved by the British Committee for Standards in Haematology

“…Misdiagnosis can be minimized by ensuring at least two concordant sets of results are obtained. Samples should not be put on ice (Bohm, et al 2006). …”

Are there sufficient standards for the in vitro hemocompatibility testing of biomaterials?

“…These guidelines recommend that blood preparation should be carried out at room temperature and variations in temperature should be avoided due to the activation of blood cells at decreased temperatures [83,84]. However, there is evidence to suggest that a temperature of 37°C, constantly kept during the blood withdrawal, storage and preparation as well as during the final assay, may reflect the in vivo situation even more appropriately [85][86][87].…”