title: Detection of alpha-thalassemias by multiplex polymerase chain reaction.
abstract: Although alpha-thalassemia is the most common genetic abnormality in the world, there is currently no routine laboratory method to definitively identify individuals who are affected. We describe a rapid and simple method that utilizes deletion-sensitive primers to amplify normal DNA sequences. Deletions involving the regions responsible for most of the alpha-thalassemia cases in the US prevent amplification with these primers. In tests with DNA isolated from small amounts (10 microL) of whole blood, the deletion-sensitive primers gave rise to the expected 248- and 375-bp (base pair) amplification products in normal individuals. These primers, along with primers designed to bind to a nonaffected control sequence from the hemoglobin beta chain, could be amplified simultaneously (multiplex polymerase chain reaction). This made it possible to detect heterozygotes for alpha-thalassemia-2 (one alpha locus deleted) by determining the ratios of the 248- and 375-bp amplification products to the product of the control sequence (268 bp). The method is rapid and simple and can be performed in a routine clinical laboratory.
authors.affiliation: Department of Pathology and Laboratory Medicine, Evanston Hospital, IL 60201-1783.
authors.forename: L J
authors.initials: LJ
authors.lastname: Bowie
authors.forename: P L
authors.initials: PL
authors.lastname: Reddy
authors.forename: M
authors.initials: M
authors.lastname: Nagabhushan
authors.forename: P
authors.initials: P
authors.lastname: Sevigny
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons; D017353:Gene Deletion; D005914:Globins / Q000235:genetics*; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017085:alpha-Thalassemia / Q000175:diagnosis* / Q000235:genetics
chemical_list: D005914:Globins; D004247:DNA