ABSTRACT
Over many years, cases of suspected a-globin chain variants were collected from different parts of the
UK. The suspicion was based on the clinical picture, high performance liquid chromatography (HPLC)
variant percentage, retention time (RT) and isoelectric focusing (IEF). DNA sequencing and the restriction
enzyme EaeI were used for definitive diagnosis. One hundred and forty-eight variants were confirmed
on one or both of the two a-globin genes (HBA2, HBA1). These cases were identified as 46
different a-globin chain variants. The most common variants were Hb J-Meerut [HBA2: c.362C>A (or
HBA1)] (10.1%) and Hb Q-India (HBA1: c.193G>C) (8.1%), followed by Hb J-Paris-I [HBA2: c.38C>A (or
HBA1)] and Hb Manitoba II (HBA1: c.309C>A) (7.4% for each). Other a variants were detected at lower
frequencies. Two novel alleles were also detected: Hb Walsgrave [a116(GH4)Glu!Val (HBA2: c.350A>T)]
and Hb Coombe Park [a127(H10)Lys!Glu (HBA2: c.382A>G)]. The majority of the ethnic origin was
Indian. The positive predictive value for a variant identification by HPLC-RT analysis was 65.9%, 41.9%
by IEF, and using both RT and IEF, the value was 72.1%. The number of variants was higher in HBA1
than in HBA2 genes and in exons 1 and 2 than in exon 3. There was no clustering of mutations in consecutive
codons. This study, the characterization of a wide spectrum of a-globin chain variants, can
facilitate the presumptive diagnosis of these variants prior to screening by a panel of amplification
refractory mutation system-polymerase chain reaction (ARMS-PCR), and a definitive diagnosis by
DNA sequencing.