Dear Editor, This paper reports a novel mutation in the coding region of the " gene. This mutation is a frameshift deletion of 4 bp (−TGGA) in the codons 20/21 of the first exon of this gene and this produces a "-thalassemia phenotype. "-Thalassemias are a heterogeneous group of autosomal recessive disorders characterized by deficient (") or absent (") "-globin chain synthesis. The large majority of the β-thalassemia defects are single nucleotide mutations that affect one of the different molecular processes involved in "-globin gene expression (transcription, pre-mRNA processing, and translation). Only a minority of "-thalassemia is produced by deletions of the structural gene. More than 200 different " gene mutations have been described so far. The largest number of "thalassemia results derived from so-called nonsense and frameshift mutations, which interrupt the normal translation of mRNA [1, http://globin.cse.psu.edu/globin/hbvar]. An asymptomatic 29-year-old man, born in Galicia (northwest of Spain), was studied following a routine blood smear analysis, which revealed microcytic and hypochromic red cells. He was referred to us as a suspected case of thalassemia. Diagnosis of β-thalassemia was performed measuring red cell indices on an automated blood cell counter (COULTER GEN-S, Coulter, Hialeah, FI, USA). The propositus exhibited a discreet anemia (Hb 12.5 g/dl and packed cell volume 38.2%); an elevated red cell count (6.39×10/l); a lower mean cell volume (59.7 fl) and reduced mean cell hemoglobin (19.6 pg). The HbA2 and HbF levels were obtained by high performance liquid chromatography (HPLC-VARIANTTM BioRad Laboratories, Hercules, CA, USA) and while HbA2 concentration was increased (5.1%) HbF level was normal (1.3%). His reticulocyte count was 0.96% (this relatively low count may be a possible consequence of bone marrow dyserythropoiesis). The subject did not show ferropenic anemia (ferritin 525 ng/ml) and the total bilirubin concentration was 1.07 mg/dl. The blood smear showed microcitosis, hypochromia, and basophilic stippling. The propositus did not show splenomegaly, hematological data, and clinical evolution was steady since 2002 (carrier of a "-thalassemia). For molecular study, DNAwas extracted from peripheral blood leukocytes using the salting out extraction procedure [2]. Initial screening, performed by real time polymerase chain reaction (RT-PCR Light Cycler, Roche) [3] and polymerase chain reaction-amplification reaction mutations system (PCR-ARMS), testing the most frequent "-thalassemia mutations in the Mediterranean area were negative [4, 5]. Finally, the characterization of the mutation was carried out by sequencing. The "-globin gene was amplified using the following primers: "1F, 5′-TCC TAA GCC AGT TGC CAG AAG-3′ [specific for the 5′ region of the "-globin gene (from nt −160 to −142)] and "1R, 5′-TCATTC GTC TGT TTC CCA TTC TAA AC-3′ (from nt +610 to +585) (all nt positions are given relative to the CAP site=nt1). The PCR product (770 bp long) was treated with the ABI PRISMTM dRhodamine Terminator Cycle Sequencing Ann Hematol (2008) 87:63–65 DOI 10.1007/s00277-007-0340-4
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