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Clinical Chemistry 44: 2381-2382, 1998;
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(Clinical Chemistry. 1998;44:2381-2382.)
© 1998 American Association for Clinical Chemistry, Inc.

Letters

Hb A2 in Subjects with Hb D

Sumitra Dash

Salmaniya Medical Centre, Bahrain, Fax 0973-279649


To the Editor:

Dr. Huisman's paper on "Combinations of ß chain abnormal hemoglobins with each other or with ß-thalassemia determinants with known mutations: influence on phenotype" (1), was of great interest to us because Bahrain has a high prevalence of hemoglobinopathies (2). Monitoring with cation-exchange HPLC (Bio-Rad Variant) was started in 1997, and 11 800 blood samples were screened in that year.

According to Dr. Huisman, when "a slowly moving variant like Hb D was present, the Hb A2 zone was often contaminated with small amounts of minor (modified) Hb D, resulting in Hb A2 values that were too high". Our results in 27 cases of Hb D trait do not agree with this. The median value for Hb A2 was 1.6% (range, 0.9–2.5%) in Hb D trait cases against a median value of 2.6% (range, 1.7–3.1%) in healthy subjects (Table 1 ). Low Hb A2 was also seen in Hb S-D disease cases.


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  		Table 1. Hb A2 concentrations in healthy subjects and HbD, HbS, and ß-thalassemia trait cases.

Our data agree with the fact that, in the presence of sickle hemoglobin, the Hb A2 peak is inappropriately high [as noted in the Bio-Rad application note and also observed by Wild and Stephens (3)].

Thus, when Hb D-B thalassemia cases are interpreted, Hb A2 values should be interpreted as in the ß-thalassemia trait. The reason for low Hb A2 in Hb D cases is unclear. A portion of Hb A2 in these cases may elute with either the A or D peak, or the mutation itself may influence the amount of delta chain, because its expression is low even in Hb S-D.


References

  1. Huisman THJ. Combinations of ß chain abnormal hemoglobins with each other or with ß-thalassemia determinants with known mutations: influence on phenotype. Clin Chem 1997;43:1850-1856. [Abstract/Free Full Text]
  2. Nadkarni KV, Al-Arrayed S, Bapat JP. Incidence of genetic disorders of haemoglobins in the hospital population of Bahrain. Bah Med Bull 1991;13:19-24.
  3. Wild BJ, Stephens AD. The use of automated HPLC to detect and quantitate haemoglobins. Clin Lab Haematol 1997;19:171-176. [ISI][Medline] [Order article via Infotrieve]

Professor Emeritus Huisman responds:

Titus H. J. Huisman

Department of Biochemistry, and Molecular Biology, Research & Education Building, Room CB-2208, Medical College of Georgia, Augusta, GA 30912-2114, Fax 706-721-3092


To the Editor:

Thank you for sending me the short note by Dr. Dash. She is probably correct that quantification of Hb A2 with her HPLC system in Hb D-Punjab-containing samples is different from that observed earlier in my laboratory; it shows how different the results can be with different columns and/or elution systems. It would be interesting to see if the Hb A2 concentrations in her Hb D-containing samples are again on the low side of normal when analyzed with other procedures (DEAE-cellulose chromatography or cellulose acetate elution procedure).




The following articles in journals at HighWire Press have cited this article:


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K. Zurbriggen, M. Schmugge, M. Schmid, S. Durka, P. Kleinert, T. Kuster, C. W. Heizmann, and H. Troxler
Analysis of Minor Hemoglobins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry
Clin. Chem., June 1, 2005; 51(6): 989 - 996.
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A. Joutovsky, J. Hadzi-Nesic, and M. A. Nardi
HPLC Retention Time as a Diagnostic Tool for Hemoglobin Variants and Hemoglobinopathies: A Study of 60000 Samples in a Clinical Diagnostic Laboratory
Clin. Chem., October 1, 2004; 50(10): 1736 - 1747.
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G. M. Clarke and T. N. Higgins
Laboratory Investigation of Hemoglobinopathies and Thalassemias: Review and Update
Clin. Chem., August 1, 2000; 46(8): 1284 - 1290.
[Abstract] [Full Text] [PDF]

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F. Cotton, B. Gulbis, V. Hansen, F. Vertongen, and S. Dash
Interference of Hemoglobin D in Hemoglobin A2 Measurement by Cation-Exchange HPLC • Dr. Dash responds:
Clin. Chem., August 1, 1999; 45(8): 1317 - 1318.
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This Article
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Right arrow Articles by Dash, S.
Right arrow Articles by Huisman, T. H. J.

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