title: Diagnosis of alpha-1-antitrypsin deficiency: An algorithm of quantification, genotyping, and phenotyping.
abstract: BACKGROUND
  Laboratory testing in suspected alpha-1-antitrypsin (A1AT) deficiency involves analysis of A1AT concentrations and identification of specific alleles by genotyping or phenotyping. The purpose of this study was to define and evaluate a strategy that provides reliable laboratory evaluation of A1AT deficiency.
  
  
  METHODS
  Samples from 512 individuals referred for A1AT phenotype analysis were analyzed by quantification, phenotype, and genotype. A1AT concentrations were measured by nephelometry. Phenotype analysis was performed by isoelectric focusing electrophoresis. The genotype assay detected the S and Z deficiency alleles by a melting curve analysis.
  
  
  RESULTS
  Of the 512 samples analyzed, 2% of the phenotype and genotype results were discordant. Among these 10 discordant results, 7 were attributed to phenotyping errors. On the basis of these data we formulated an algorithm, according to which we analyzed samples by genotyping and quantification assays, with a reflex to phenotyping when the genotype and quantification results were not concordant. Retrospective analyses demonstrated that 4% of samples submitted for genotype and quantitative analysis were reflexed to phenotyping. Of the reflexed samples, phenotyping confirmed the genotype result in 85% of cases. In the remaining 15%, phenotyping provided further information, including identifying rare deficiency alleles and suggesting the presence of a null allele, and allowed for a more definitive interpretation of the genotype result.
  
  
  CONCLUSIONS
  The combination of genotyping and quantification, with a reflex to phenotyping, is the optimal strategy for the laboratory evaluation of A1AT deficiency.
authors.affiliation: Division of Clinical Biochemistry & Immunology, Department of Laboratory Medicine & Pathology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
authors.forename: Melissa R
authors.initials: MR
authors.lastname: Snyder
authors.forename: Jerry A
authors.initials: JA
authors.lastname: Katzmann
authors.forename: Malinda L
authors.initials: ML
authors.lastname: Butz
authors.forename: Carmen
authors.initials: C
authors.lastname: Wiley
authors.forename: Ping
authors.initials: P
authors.lastname: Yang
authors.forename: D Brian
authors.initials: DB
authors.lastname: Dawson
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Halling
authors.forename: W Edward
authors.initials: WE
authors.lastname: Highsmith
authors.forename: Stephen N
authors.initials: SN
authors.lastname: Thibodeau
mesh_terms: D000465:Algorithms; D042241:Early Diagnosis; D005838:Genotype; D006801:Humans; D009391:Nephelometry and Turbidimetry; D010641:Phenotype; D012189:Retrospective Studies; D000515:alpha 1-Antitrypsin / Q000235:genetics*; D019896:alpha 1-Antitrypsin Deficiency / Q000175:diagnosis*
chemical_list: D000515:alpha 1-Antitrypsin