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.
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.
Papassotiriou
I
I
Department of Clinical Biochemistry, Aghia Sophia, Children's Hospital, Athens, Greece. pan-gr@ath.forthnet.gr
Traeger-Synodinos
J
J
Vlachou
C
C
Karagiorga
M
M
Metaxotou
A
A
Kanavakis
E
E
Stamoulakatou
A
A
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
D006455:Hemoglobins, Abnormal; D005914:Globins; D006447:Hemoglobin H; C030448:hemoglobin Bart's