Screening for Intermediate and Severe Forms of Thalassaemia in Discarded Red Blood Cells: Optimization and Feasibility
2011
SUMMARY Detection and quantification of Hb subtypes of human blood is integral to presumptive identification of
thalassaemias. It has been used in
neonatal screeningof
thalassaemiaand Hb variants. The use of discarded
red blood cellsfollowing processing of the
cord bloodfor stem cells provides readily available diagnostic material for
thalassaemiascreening. In this study, we determined the range of Hb subtypes in 195 consecutive
cord bloodsamples collected for
cord bloodbanking. Thecord blood samples' analysed were those of the remaining
red blood cellsafter the
cord bloodwas processed for stem cell storage. Quantification of Hb subtypes by high performance liquid chromatography (HPLC) was done on BioRad Variant II Hb testing system. Only 73 (36.5%) of the samples could be analyzed neat without dilution. With a 1:300 dilution with wash solution the acceptable area as recommended by the manufacturer for reading of a C-gram within the 1 to 3 million ranges were achieved in all. Eighteen (9%) 12 showed classical Hb Barts (γ4) prerun peaks were confirmed by Sebia Hydrasys automated Hb gel electrophoresis and quantified by Sebia Capillarys 2 capillary electrophoresis. Only 1 (0.5%) was presumptively identified with HbH disease. Due to the limited number of samples no beta-
thalassaemia major, Hb E
beta-thalassaemiaand Hb Barts
hydrops fetaliswere found. The HPLC assay was possible at a cost US$ 5 per sample and a turnover time of 10 samples per hour without technical difficulties. This study reports an effective and valuable protocol for
thalassaemiascreening in
red blood cellswhich would otherwise be discarded during
cord bloodprocessing.
Cord bloodwith severe and intermediate forms of
thalassaemiacan be preselected and not stored.
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