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Título

Higher stability of basophil activation test read-out simplifies assay performance and sample logistics.

Resumo

Background: Basophil Activation Tests (BAT) have gained increasing importance in the field of allergy diagnostics, supported by growing scientific evidence for the higher accuracy and clinical relevance over other allergy tests. BAT is a functional allergy test based on viable basophils that are detected by the low-mid throughput flow cytometry technology. Therefore, the appropriate conditions for specimen storage and the management of acquisition of processed samples are crucial for the widely use of BAT testing.
Objective: To improve logistics, practicability and time management of BAT testing, the novel version of the BAT including a stabilizing agent, has been investigated for stimulated and processed basophils.
Methods: Two separated studies were performed using whole blood from four normal blood donors. First, the specimen stability of unprocessed EDTA whole blood was assessed by storing fresh blood at different temperatures (2-8°C and 28°C) for 0 to 4 days before performing the BAT assay. Second, the stability of processed samples was assessed at different temperatures (2-8°C and 28°C) and measured at different time points (0 to 10 days) after cell stimulation and fixation.
Results: Unprocessed EDTA whole blood samples of all four donors were stable for 48 hours when stored at 2-8°C and for 24 hours stored at 28°C before basophil activation with at least 80% recovery. Processed and fixed basophils remained stable for flow cytometry acquisition for at least 10 days at 2-8°C (min. 80% recovery), while for 28°C, 48 hours stability could be shown.

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