title: Polymerase chain reaction for detection of the alpha-1-antitrypsin Z allele in chronic liver disease.
abstract: The genetic locus for alpha-1-antitrypsin (alpha-AT) is highly polymorphic, but all protein variants are encoded by a single locus on chromosome 14. Periportal hepatocyte granules are described in association with chronic liver disease and the Z variant. A Z-specific point mutation in exon V of the alpha-AT gene, converting amino acid 342 from Glu to Lys, is thought to be responsible for the hepatocyte accumulation. We describe the use of the polymerase chain reaction (PCR) to amplify exon V of the alpha-AT gene and subsequent detection of the wild-type M- and Z-specific sequences by hybridisation to 32P-labelled-allele-specific oligonucleotides. We applied this technique to leucocyte DNA from 37 patients with suspected chronic liver disease, 25 of whom had hepatocyte alpha-AT inclusion granules on liver biopsy. All 25 were homozygous or heterozygous for the Z allele. One patient, phenotyped as PiS, was found to be PiSZ and another phenotyped as PiZ (presumed homozygous), was found to be a Z heterozygote. No Z allele was detected in any of the twelve patients without alpha-AT inclusion granules. This sensitive PCR technique could be used to assess the relative risk of chronic liver disease in PiZ heterozygotes and to determine whether individuals without the Z amino acid 342 substitution can developed periportal alpha-AT granules.
authors.affiliation: Department of Medicine, University of Newcastle Upon Tyne, United Kingdom.
authors.forename: A M
authors.initials: AM
authors.lastname: Brind
authors.forename: I
authors.initials: I
authors.lastname: McIntosh
authors.forename: D J
authors.initials: DJ
authors.lastname: Brock
authors.forename: O F
authors.initials: OF
authors.lastname: James
authors.forename: M F
authors.initials: MF
authors.lastname: Bassendine
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002908:Chronic Disease; D005091:Exons; D005838:Genotype; D006801:Humans; D015151:Immunoblotting; D007525:Isoelectric Focusing; D008107:Liver Diseases / Q000235:genetics*; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D000515:alpha 1-Antitrypsin / Q000235:genetics*
chemical_list: D000515:alpha 1-Antitrypsin