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Published on: September 16, 2012
Long-term stabilized blood samples as controls for flow cytometric HLA-B27 screening: a feasibility study
Wilfried H B M Levering1, Henk Wind, Vivian Granger
1Laboratory for Histocompatibility and Immunogenetics, Sanquin Blood Bank South West Region, Rotterdam, The Netherlands. w.levering@sanquin.nl
Cytometry. Part B, Clinical Cytometry
|January 18, 2008
Summary
Long-term stabilized blood samples can serve as controls for flow cytometric HLA-B27 screening and external quality assessment. While feasible for quality control, their variability currently limits reagent validation and requires extensive quality checks.
Area of Science:
- Immunology
- Hematology
- Clinical Chemistry
Background:
- Long-term stabilized blood samples offer potential as controls for flow cytometric HLA-B27 screening.
- These samples could also be utilized in external quality assessment (EQA) schemes.
Purpose of the Study:
- To evaluate the utility of long-term stabilized whole blood specimens for HLA-B27 screening.
- To assess the stability and performance of these samples over time.
Main Methods:
- Peripheral blood from nine donors was stabilized using Trans-FIX.
- Stabilized and fresh samples were tested with anti-HLA-B27 monoclonal antibodies over 12 months.
- Flow cytometry parameters (FSC, SSC, fluorescence) were analyzed, and a pilot EQA study was conducted.
Main Results:
- Lymphocyte scatter characteristics (FSC, SSC) remained largely unaffected.
- Stabilized samples showed increased background and decreased specific fluorescence compared to fresh samples.
- Discrimination between HLA-B27 positive and negative samples was feasible, but concordance in the pilot EQA was lower due to sample variability.
Conclusions:
- Long-term stabilized whole blood is suitable for HLA-B27 positive/negative controls in daily and longitudinal quality control.
- Stabilized samples show promise for EQA but are not yet suitable for reagent validation.
- Rigorous quality control is essential before using stabilized samples in multicenter studies.
