Efficacy of a modified improved technique for detecting red cell haemoglobin H inclusions

Abstract

Summary. Alpha‐thalassaemia is a common disease in Taiwan. A feature useful in diagnosis is the excess of of β‐chains that result from impaired alpha‐chain production. These excess chains assemble into β4 tetramers (i.e. Hb H) which can be detected by its rapid anodal migration on alkaline electrophoretic media as seen in Hb H disease. However, this technique cannot, and conventional Hb H inclusion staining rarely can visualize α‐thalassaemic traits due to small quantities of Hb H formed in these patients. The staining for Hb H inclusion bodies uses brilliant cresyl blue (BCB) or methylene blue (MB) as an oxidant to denature Hb H as intracellular inclusions. We have improved the technique of Jones (who modified the original method in order to obtain enriched young red cells) by prolonging the incubation time from 30 min to 3 h. The sensitivity of this modified improved method was 91% for detecting obligatory α‐thalassaemic traits as shown in table. No false positive results were seen in β‐thalassaemia or in others. We believe this can be used as a confirmatory test in heterozygous α1‐thalassaemia and homozygous β2‐thalassaemia.

Fine-tiling array CGH to improve diagnostics for α- and β-thalassemia rearrangements

“…Biochemical analysis was performed on HPLC VARIANT II (Bio‐Rad Laboratories, Hercules, CA) using the β‐Thalassaemia Short Programme, and on the Capillarys capillary electrophoresis (CE) device (Sebia, Evry, France). Presence of cellular inclusions of hemoglobin H (Hb H) was determined after incubation with brilliant cresyl blue [Lin et al, 1990], in cases suspected of having α 0 ‐thalassemia. Genomic DNA was isolated using the Qiagen AutoPure LS Genomic DNA Purification System (Gentra Systems, Minneapolis, MN).…”

Alternative method of smearing samples on slides facilitate visualization of hemoglobin H inclusion bodies in alpha thalassemia

“…3 This simple test is useful for the diagnosis of alpha thalassemia, however, the identification of inclusion bodies is laborious and the results are highly observer dependent. 4,5 In an attempt to improve the identification of inclusion bodies, we tested an alternative method of smearing samples on slides.…”