title: Rapid and accurate quantitation of Hb Bart's and Hb H using weak cation exchange high performance liquid chromatography: correlation with the alpha-thalassemia genotype.
abstract: The Bio-Rad Variant Hemoglobin testing system is an automated high performance liquid chromatography analyzer marketed with a beta-thalassemia short program to quantify Hbs F and A2, and assist in detecting Hbs A, S, D, C, and E. Although the two hemoglobins present in Hb H disease, Hb Bart's and Hb H, are separated by the system, they are not quantitated. In this study we modified the beta-thalassemia short program in order to facilitate quantitation of Hb Bart's and Hb H. Blood samples from 60 patients with Hb H disease, with various underlying genotypes, were studied. Analyses were performed on the day of blood collection or on hemolysates stored at -80 degrees C in cyanide or carbomonoxy forms. The mean sum of Hb Bart's and Hb H levels in all patients was found to be 12% (range 1.8-35%). Patients with nondeletional mutations (or association of alpha(0) deletion and nondeletional mutations) had notably higher Hb Bart's and Hb H levels when compared to patients with deletional genotypes.
authors.affiliation: Department of Clinical Biochemistry, Aghia Sophia, Children's Hospital, Athens, Greece. pan-gr@ath.forthnet.gr
authors.forename: I
authors.initials: I
authors.lastname: Papassotiriou
authors.forename: J
authors.initials: J
authors.lastname: Traeger-Synodinos
authors.forename: C
authors.initials: C
authors.lastname: Vlachou
authors.forename: M
authors.initials: M
authors.lastname: Karagiorga
authors.forename: A
authors.initials: A
authors.lastname: Metaxotou
authors.forename: E
authors.initials: E
authors.lastname: Kanavakis
authors.forename: A
authors.initials: A
authors.lastname: Stamoulakatou
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005838:Genotype; D005914:Globins / Q000235:genetics; D006447:Hemoglobin H / Q000032:analysis*; D006455:Hemoglobins, Abnormal / Q000032:analysis*; D006801:Humans; D007231:Infant, Newborn; D007525:Isoelectric Focusing / Q000379:methods; D009154:Mutation; D015203:Reproducibility of Results; D017085:alpha-Thalassemia / Q000235:genetics / Q000378:metabolism
chemical_list: D006455:Hemoglobins, Abnormal; D005914:Globins; D006447:Hemoglobin H; C030448:hemoglobin Bart's