Preanalytic variables of thrombin generation: towards a standard procedure and validation of the method
| Autor: | Rinske Loeffen, Marie-Claire Kleinegris, Henri M. H. Spronk, Diane Fens, H. ten Cate, Sarah T.B.G. Loubele, R. van Oerle, P. H. M. Pluijmen |
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| Přispěvatelé: | Promovendi CD, Biochemie, MUMC+: DA CDL Analytisch cluster 1K (9), Interne Geneeskunde, RS: CARIM School for Cardiovascular Diseases |
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
validation
medicine.medical_specialty Venipuncture business.industry preanalytic variables Thrombin Centrifugation Hematology medicine.disease Thrombin generation Hemolysis Standard procedure Surgery Tissue factor Reference sample collection tube Reference Values thrombin generation Butterfly needles medicine Humans business Whole blood Biomedical engineering |
| Zdroj: | Journal of Thrombosis and Haemostasis, 10(12), 2544-2554. Wiley |
| ISSN: | 1538-7836 1538-7933 |
| DOI: | 10.1111/jth.12012 |
| Popis: | Summary. Background: Thrombin generation assays are sensitive methods for assessment of the overall clotting potential of plasma, but, despite their common use in thrombosis research, standardization of preanalytic conditions is lacking. In order to set up a standardized protocol, we analyzed different preanalytic variables and validated the calibrated automated thrombogram method. Methods and Results: Thrombin generation was assessed with 0, 1 and 5 pm tissue factor (TF). Variations in thrombin generation were mostly attributable to the type of collection tube, mainly because of variations in contact activation. The collection tube also determined the influence of other preanalytic variables on thrombin generation, e.g. the need for a discard tube, the storage of whole blood, and the centrifugation method. Regarding the collection system, blood drawn through intravenous catheters or butterfly needles showed significantly more hemolysis than blood obtained with venipuncture using conventional needles. The results showed that a discard tube is still needed for thrombin generation measurements. After blood collection, whole blood is best centrifuged immediately, to prevent activation or degradation of coagulation proteins, and a second centrifugation step at 10 000 × g is recommended. After thawing, plasma is best analyzed immediately, as storage resulted in thrombin generation results outside the 10% range of the reference sample. On the basis of these results, we set up an in-house standardized protocol, which was used for validation, resulting in coefficients of variations of |
| Databáze: | OpenAIRE |
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