Nonradioactive quantification of low concentrations of hemoglobin A by HPLC for midtrimester prenatal diagnosis of β-thalassemia

R. Maiavacca, S. Tedeschi, A. Mosca, S. Calmi, P. De Leonardis, C. Todaro, G. Zuliani, M. Buscaglia, A. Cantu-Rajnoldi

Research output: Contribution to journalArticle

Abstract

The usual methods for prenatal diagnosis of β-thalassemia and other hemoglobinopathies by assay of fetal blood erythrocytes are either complex (analysis of globin chains synthesis by carboxymethylcellulose chromatography) or only semiquantitative [isoelectric focusing of hemoglobin (Hb)]. To further simplify the diagnostic procedure and to obtain quantitative data, we measured the small concentrations of Hb A in fetal erythrocytes by using a high-pressure liquid chromatography (HPLC) instrument (DIAMAT-TM; Bio-Rad) equipped with the new column proposed for measuring Hb A 2. We analyzed 212 uncontaminated fetal blood samples obtained by cordocentesis between the 18th and 22nd weeks of pregnancy, using the HPLC procedure, and compared the results with those obtained by the above-named methods. The Hb A values obtained ranged between 0% and 8.5%; they were ≤1.8% in 44 fetuses affected by homozygous β-thalassemia and >2.5% in 168 unaffected fetuses. The method was simple, rapid, and reproducible (CV 3.2%) and there was good correlation between Hb A concentrations determined by HPLC and the β/γ ratio determined by carboxymethylcellulose chromatography (r = 0.7687; P

Original languageEnglish
Pages (from-to)1906-1908
Number of pages3
JournalClinical Chemistry
Volume38
Issue number9
Publication statusPublished - 1992

Keywords

  • erythrocytes
  • fetal status
  • hemoglobin variants
  • hemoglobinopathies

ASJC Scopus subject areas

  • Clinical Biochemistry

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    Maiavacca, R., Tedeschi, S., Mosca, A., Calmi, S., De Leonardis, P., Todaro, C., Zuliani, G., Buscaglia, M., & Cantu-Rajnoldi, A. (1992). Nonradioactive quantification of low concentrations of hemoglobin A by HPLC for midtrimester prenatal diagnosis of β-thalassemia. Clinical Chemistry, 38(9), 1906-1908.