Japanese collaborative study to assess inter-laboratory variation in factor VII activity assays.

BACKGROUND: The clinical phenotype manifest by patients with factor VII (FVII) deficiency correlates poorly with that predicted by laboratory tests. Despite its importance, there are no data on the variability of inter-laboratory determinations of low to very low plasma FVII activity (FVII:C). METHODS: We distributed three FVII-deficient plasma samples, prepared by immunoaffinity chromatography, to 58 laboratories in Japan. All samples were assayed using standardized reference plasma as a calibrator. Recombinant thromboplastin was also supplied as a common reagent. RESULTS: In the case of sample A, which had a very low FVII:C, the use of standardized reference plasma and thromboplastin, lowered the variability of inter-laboratory measurements, when compared with the variability observed when samples were assayed using the respective laboratory's routine method. CONCLUSIONS: The data obtained indicated that results for samples with a very low FVII:C were greatly influenced by the number of plasma dilutions used in constructing a standard activity curve, and also by the type of calibrator and thromboplastin. Such variability was not seen for samples with moderate FVII:C. We conclude that it is necessary to develop a more sensitive and accurate FVII:C measurement system for the diagnosis and treatment of FVII deficiency.

Japanese collaborative study for fibrinogen assay: variability of the fibrinogen assay between different laboratories does not improve when a common calibrator is used.

Several national and local external quality assurance schemes have been developed to improve the plasma fibrinogen assay in Japan over the past 30 years. Now most commercial calibrant plasma may be calibrated against an International Standard preparation, in order to achieve agreement of results obtained by different laboratories. However, we have never achieved satisfactory results, according to an external quality control survey regarding the fibrinogen assay. Therefore, we distributed two kinds of fibrinogen standards to be used as common calibrators, along with three plasma samples, among 183 general laboratories in Japan. The results of this collaborative study showed that the assigned value for the commercially available calibrators remained problematic. Furthermore, it was concluded that the between-laboratory variability could not be improved beyond a certain degree of standardization, even if a common calibrator was used for the Clauss-derived assay carried out by an automatic coagulometer.

Use of theophylline in the investigation of pseudothrombocytopenia induced by edetic acid (EDTA-2K).

In automated cell counting of edetic acid (EDTA-2K) anticoagulated blood, thrombocytopenia is occasionally seen which bears no relation to any underlying disease. In this study a heparin and soluble theophylline mixture was used to measure accurately platelet numbers in patients with such pseudothrombocytopenia. In four normal volunteers, a theophylline concentration of more than 7 mg/ml produced no significant difference in platelet numbers between theophylline and heparin and EDTA-2K anticoagulated bloods. When blood treated with EDTA-2K was used in seven patients with pseudothrombocytopenia, falsely low platelet counts were observed in three patients immediately after sampling; in blood treated with theophylline, white cell and platelet counts remained unchanged for up to six hours after sampling. Microscopical examination of the EDTA-2K anticoagulated blood showed massive platelet clumping, but no aggregates were seen in theophylline anticoagulated blood. It is concluded that theophylline can be useful in the investigation of pseudothrombocytopenia when an automated cell counter is used.