Hemoglobin H identification by high-performance liquid chromatography in confirmed hemoglobin H disease.

Abstract

INTRODUCTION:

Among hemoglobin (Hb) H disease cases diagnosed by DNA testing in our hemoglobinopathy laboratory, we have noted instances of unreported Hb H from high-performance liquid chromatography (HPLC) results of referring laboratories.

METHODS:

To characterize these issues, we identified all cases of genotypic Hb H disease diagnosed in our laboratory. HPLC chromatograms were reviewed to determine the presence and retention time of the Hb H peak.

RESULTS:

Hemoglobin H was not reported in 24.2% of patients (23 of 95) with genotypic Hb H disease. The characteristic prerun peak of Hb H was present on review of all eight Variant or Variant II β-thalassemia short-program chromatograms. Elevated Hb F (≥3%) was reported in 14 cases. The Hb H peak was found in the Hb F window in 11 dual program cases. The incorrect identification of Hb H as elevated Hb F resulted in two testing referrals for 'δβ-thalassemia'.

CONCLUSIONS:

Hemoglobin H may go unreported due to failure to examine for or recognize its peak on Variant or Variant II β-thalassemia short-program chromatograms. Elution of Hb H in the Hb F window resulted in misidentification of Hb H for Hb F and may indicate a Variant II HbA2 /HbA1C program software error. Our findings highlight the need for careful chromatogram inspection and clinical correlation in the diagnosis of Hb H disease.

• Authors: E Turley, A McFarlane, L Halchuk, M Verhovsek
• Journal: International journal of laboratory hematology (Int J Lab Hematol) Vol. 37 Issue 5 Pg. 668-72 (Oct 2015) ISSN: 1751-553X [Electronic] England
• PMID: 25989219 (Publication Type: Journal Article)
• Copyright: © 2015 John Wiley & Sons Ltd.

Chemical References

• alpha-Globins
• Fetal Hemoglobin
• Hemoglobin H

Topics

• Adult
• Chromatography, High Pressure Liquid (methods)
• Erythrocyte Indices
• Female
• Fetal Hemoglobin (chemistry)
• Genotype
• Hemoglobin H (chemistry, genetics)
• Humans
• Male
• Middle Aged
• Young Adult
• alpha-Globins (chemistry, genetics)
• alpha-Thalassemia (blood, diagnosis, genetics)