title: HPLC determination of hemoglobins to establish reference values with the aid of statistics and informatics.
abstract: The purpose of the present study was to establish reference values for hemoglobins (Hb) using HPLC, in samples containing normal Hb (AA), sickle cell trait without alpha-thalassemia (AS), sickle cell trait with alpha-thalassemia (ASH), sickle cell anemia (SS), and Hb SC disease (SC). The blood samples were analyzed by electrophoresis, HPLC and molecular procedures. The Hb A2 mean was 4.30 +/- 0.44% in AS, 4.18 +/- 0.42% in ASH, 3.90 +/- 1.14% in SS, and 4.39 +/- 0.35% in SC. They were similar, but above the normal range. Between the AS and ASH groups, only the amount of Hb S was higher in the AS group. The Hb S mean in the AS group was 38.54 +/- 3.01% and in the ASH it was 36.54 +/- 3.76%. In the qualitative analysis, using FastMap, distinct groups were seen: AA and SS located at opposite extremes, AS and ASH with overlapping values and intermediate distribution, SC between heterozygotes and the SS group. Hb S was confirmed by allele-specific polymerase chain reaction. The Hb values established will be available for use as a reference for the Brazilian population, drawing attention to the increased levels of Hb A2, which should be considered with caution to prevent incorrect diagnoses.
authors.affiliation: Laboratório de Hemoglobinas e Genética das Doenças Hematológicas, Departamento de Biologia, IBILCE/UNESP, São José do Rio Preto, SP, Brasil. luondei@yahoo.com.br
authors.forename: L S
authors.initials: LS
authors.lastname: Ondei
authors.forename: P J A
authors.initials: PJ
authors.lastname: Zamaro
authors.forename: P H
authors.initials: PH
authors.lastname: Mangonaro
authors.forename: C R
authors.initials: CR
authors.lastname: Valêncio
authors.forename: C R
authors.initials: CR
authors.lastname: Bonini-Domingos
mesh_terms: D001938:Brazil; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019295:Computational Biology / Q000379:methods; D004586:Electrophoresis / Q000379:methods; D005821:Genetic Techniques; D006454:Hemoglobins / Q000032:analysis / Q000737:chemistry*; D006579:Heterozygote; D006801:Humans; D015233:Models, Statistical; D009154:Mutation; D010641:Phenotype; D012016:Reference Values; D012805:Sickle Cell Trait / Q000097:blood; D017085:alpha-Thalassemia / Q000097:blood
chemical_list: D006454:Hemoglobins