Comparison of HPLC results of patients with hemoglobin D with DNA sequence analysis: Detection of compound heterozygosity HbD/β-Thalassemia traits

Authors

DOI:

https://doi.org/10.5281/zenodo.14183773

Keywords:

DNA analysis, Hemoglobin D, Hemoglobin variant analysis, HPLC, β-Talassemia

Abstract

Objective: In our study, we aimed to compare the results of patients with near-total hemoglobin D variants detected by the HPLC method with DNA sequence analysis.

Materials and methods: In our laboratory, hemoglobin variant analysis was performed with an HPLC analyzer (Adams A1C HA8180T, Arkray, Inc., Kyoto, Japan) from EDTA-tubed whole blood samples of three male patients. PCR-DNA Sanger sequence analysis was then performed on these samples for a definitive diagnosis.

Results: We found that three male patients had hemoglobin D variants close to the total by HPLC method (HbD levels are 97.92%, 93.2%, and 97.6%, and HbA levels are 0% for all patients, respectively). In two patients, we did not detect hemoglobin A2 levels by HPLC, while in one patient, we detected <3.5% HbA2. According to the results of PCR-DNA sequence analysis, we found that all three patients had heterozygous Hemoglobin D, characterized by the p.Glu121Gln (c.364G>C) mutation. In addition, the patients had c.92 +5G>C, IVS2-1G>A (c.315+1G>A), and p.Lys9Valfs*14(c.25_26del) mutations, which were pathologically identified and consistent with heterozygous β-thalassemia carriage. Thus, we found that three patients had compound heterozygous HbD/β-thalassemia.

Conclusion: The presence of hemoglobin D is close to total hemoglobin D in percentage, according to the HPLC result, but does not always indicate homozygous hemoglobin DD. In addition, β-thalassemia carriage may be missed from the laboratory results of those who are carriers of the hemoglobin D variant and have nonincreasing hemoglobin A2 levels. In these patients, it should be clarified whether they are β-thalassemia carriers by genetic methods. In this way, the birth of babies with β-thalassemia major can be prevented.

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Published

2024-09-30

How to Cite

1.
Aktas A, Yilmazsamli L, Ongen Ipek B, Vurgun E, Sahin M, Dikker O, Dag H. Comparison of HPLC results of patients with hemoglobin D with DNA sequence analysis: Detection of compound heterozygosity HbD/β-Thalassemia traits. J Clin Trials Exp Investig [Internet]. 2024 Sep. 30 [cited 2024 Dec. 13];3(3):85-90. Available from: https://jctei.com/index.php/jctei/article/view/137