title: Simultaneous detection of Hb constant spring (α142, TAA>CAA, α2) and the α2 IVS-I donor site (-TGAGG) deletion by a simple polymerase chain reaction-based method in Iran.
abstract: Hb Constant Spring (Hb CS, codon 142, TAA>CAA, α2) (HBA2:c.427T>C) and α2 IVS-I donor site (GAGGTGAGG>GAGG - - - - -) (HBA2:c.95+2_95+6delTGAGG) are nondeletional α-thalassemia (α-thal) mutations found all over the world. Identification of α-thal genotypes in at-risk couples for severe anemia or in highly heterogeneous populations requires rapid, accurate and cost-effective genotyping methods. In this study, a pair of primers were used to specifically amplify an 883 bp fragment from the α2-globin gene in order to simultaneously identify these two mutations by a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method. We determined the genotypic frequencies of Hb CS and the α2 IVS-I donor site mutations after amplification and enzymatic digestion with Tru9I in 238 northern Iranian samples referred for α-thal testing. Hb CS and the α2 IVS-I donor site mutations accounted for 21 (8.8%) and 29 (12.2%) of the nondeletional cases. This genotyping assay has proven to be a rapid, reliable and useful diagnostic tool for simultaneous detection of these two anomalies for genetic counseling or further prenatal diagnosis.
authors.affiliation: Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Islamic Republic of Iran. halehakhavan@yahoo.com
authors.forename: Haleh
authors.initials: H
authors.lastname: Akhavan-Niaki
authors.forename: Ali
authors.initials: A
authors.lastname: Banihashemi
authors.forename: Amrollah
authors.initials: A
authors.lastname: Mostafazadeh
authors.forename: Vahid
authors.initials: V
authors.lastname: Kholghi Oskooei
authors.forename: Mandana
authors.initials: M
authors.lastname: Azizi
authors.forename: Reza
authors.initials: R
authors.lastname: Youssefi Kamangar
authors.forename: Maryam Mitra
authors.initials: MM
authors.lastname: Elmi
mesh_terms: D001483:Base Sequence; D003062:Codon; D004262:DNA Restriction Enzymes / Q000235:genetics / Q000378:metabolism; D005091:Exons; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D060005:Genotyping Techniques; D006455:Hemoglobins, Abnormal / Q000235:genetics*; D006801:Humans; D007438:Introns; D007492:Iran; D008297:Male; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017384:Sequence Deletion; D055542:alpha-Globins / Q000235:genetics*; D017085:alpha-Thalassemia / Q000175:diagnosis / Q000235:genetics
chemical_list: D003062:Codon; D006455:Hemoglobins, Abnormal; D055542:alpha-Globins; C035541:Hemoglobin Constant Spring; D004262:DNA Restriction Enzymes