Hossein Karami's research while affiliated with Mazandaran University of Medical Sciences and other places

Background: Alpha thalassemia is one of the most common human genetic abnormalities. More than 400 different variations of the α-globin protein have been introduced, most of which are not associated with noticeable clinical manifestations. The identification of all variants of Hb in different regions helps in acquiring comprehensive knowledge concerning thalassemia disease, and it can be used in preventive programs as well as prenatal diagnosis (PND). Aims: In the present study, we describe a new α1 gene mutation that leads to a frameshift after codon 83. Methods: As a plan for a national screening program of thalassemia, routine cell blood count (CBC) and Hb capillary electrophoresis tests were applied. After taking written informed consent, genomic DNA was extracted, and, for identifying common Mediterranean α-Globin gene deletion, multiplex Gap-PCR was performed; for detecting other mutations on α- and β-Globin genes, a DNA sequencing method was used. Results: The results of CBC and capillary electrophoresis tests showed microcytosis in a female subject. The sequencing of the α-Globin gene showed that the case is heterozygote for a single-nucleotide deletion at codon 83 of the α1-Globin Gene. We named this mutation Hb Adrian (α1: c.251–T), which is a novel mutation. The mentioned mutation was also detected in the subject’s mother. Conclusions: The introduced mutation (Hb Adrian) leads to a frameshift change that produces a protein with 100 amino acids, which in comparison to a normal α-chain is shorter, and its amino acids are altered after codon 83. This hemoglobin is undetectable via the use of electrophoresis. Although no major hematological abnormalities were observed in the carriers, Hb Adrian should be considered in screening programs to help prevent Hb H disease in high-risk couples.

This is a report of a couple with abnormal hematological indices who were investigated for α & β-thalassemia mutations. Based on CBC and capillary hemoglobin electrophoresis results, the male and female subjects were β & α-thalassemia carriers, respectively. Multiplex-Gap-PCR and Sanger sequencing techniques were used for the identification of mutations on α and β-globin genes. The DNA test showed the presence of c.315 + 1 G > A mutation on β-globin gene of male subject while the female case had – MED double gene deletion and c.427T > C mutation on α-globin and, interestingly, she was also a carrier for c.315 + 1 G > A mutation on β-globin gene. Cases with the coinheritance of heterozygous β0-thalassemia with one functional α-globin gene have normal HbA2 levels that may lead to their being misdiagnosed as β-thalassemia carriers, especially in premarital screening programs for thalassemia. Therefore, β-globin gene sequencing is recommended in cases with normal Hb electrophoresis and reduced hematological indices in premarital screening programs for thalassemia, especially in regions with a high frequency of β-globin mutations, in order to identify all the β-thalassemia carriers.

Objective To date, more than 400 alpha chain variants or point mutations in the α-globin genes that lead to single amino acid substitutions have been described. Here, we state a rare alpha-globin gene variant Hb Fontainebleau [a21 (B2) Ala>Pro] identified in heterozygote state in two families from the north of Iran during premarital thalassemia screening program. Material Two first cousin couples with reduced hematological indices compatible with α-thalassemia were referred to Lab for premarital screening of thalassemia. At first CBC and capillary electrophoresis tests were applied. For molecular investigation genomic DNA was isolated from peripheral blood samples. Then, multiplex-Gap PCR and Sanger sequencing techniques were applied to discover common α- globin deletions and probable point mutations, respectively. Results In the capillary electrophoresis results, one unknown peak was observed to the left of Hb A. Sanger sequencing results revealed that all cases are heterozygote for Hb Fontainebleau (HBA2: c.64G4C, p.Ala21Pro). Family history analysis has shown that all presented cases have belonged to the families which are originally from India. Conclusion The presence of Hb Fontainebleau in Mazandaran province and in subjects with Indian origin may indicate the founder effect phenomenon. DNA analysis of subjects with reduced hematological indices is recommended for finding common and rare mutations to predict childbirth with thalassemia in premarital screening programs.