Accidental detection of clinically silent compound heterozygous Hb D Punjab/Hb Q India while analyzing HbA1c by high performance liquid chromatography
DOI:
https://doi.org/10.18203/2320-6012.ijrms20214432Keywords:
HbA1c, Hb D Punjab, Hb Q India, Compound heterozygous, HPLC, Capillary electrophoresisAbstract
HbA1c is routinely prescribed investigations for diagnosing and monitoring diabetes and high-performance liquid chromatography (HPLC) is preferred method which is also able to identify presence of hemoglobin variant. A case was encountered where presence of variant hemoglobin was indicated. On further investigation with three different instruments, diagnosis of compound heterozygous Hb D Punjab/Hb Q India was made. The chromatogram on Bio-Rad D10 showed Hb D Punjab (ααββHbD Punjab)-29.89% at 3.96 minutes retention time (RT), Hb Q India (ααHbQ Indiaββ) -9.5% with 4.45 minutes RT, hybrid of HbQ India/Hb D Punjab (ααHbQ IndiaββHbD Punjab)-6% with 4.66 minutes RT, Hb A2 (ααδδ) was 2.5% and Hb A (ααββ) was 52.2%. Analysis done on Bio-Rad variant V-II confirmed these findings. Analysis done on Sebia capillary electrophoresis revealed major peak of 50.9% in zone 9/Z(A) constituted by Hb A, second peak of 39.8% in zone 6/Z(D) constituted by co-elution of Hb D and Hb Q India, third peak of 8.8% in zone 3-4/Z(A2-C) constituted by co-elution of Hb A2 and hybrid of Hb D Punjab/Hb Q India and a fourth peak of 0.5% in zone 1 representing Hb A2HbQ India (ααHbQ Indiaδδ). Ideally variants detected while analyzing HbA1c should be further investigated for confirmation and result of which should be shared, discussed and the patient should be encouraged for screening of available family members for relevant variant hemoglobin. Combination of cation exchange HPLC and capillary electrophoresis in this case was sufficient to arrive at conclusion.
References
Nasir NM, Thevarajah M, Yean CY. Haemoglobin variants detected by hemoglobin A1c (HbA1c) analysis and the effects on HbA1c measurements. Int J Diabetes Dev Ctries. 2010;30(2):86-90.
Behan KJ, Storey NM, Lee HK. Reporting variant hemoglobins discovered during hemoglobin A1c analysis- common practices in clinical laboratories. Clin Chim Acta. 2009;406:124-8.
Dangi CBS, Kaur M, Banerjee N, Angastiniotis M, Eleftheriou A, Singh M et al. Burden of Haemoglobinopathies in Madhya Pradesh, Central India. Int J Biotechnol. 2013;111:224-42.
WHO-Executive Board EB118/5. Prevalence of Haemoglobinopathies. 118th Session Report by the Secretariat on Thalassaemia and other haemoglobinopathies. 2006;1-8.
Desai D, Dhanani HJ, Shah M, Dayal N, Kapoor A, Yeluri S. Homozygous Hemoglobin D Disease: A Case Report. Internet J Pathol. 2003;3(1):1-4 .
Shanthala Devi AM, Rameshkumar K, Sitalakshmi S. Hb D: A Not So Rare Hemoglobinopathy. Indian J Hematol Blood Transfus. 2016;32(1):294-8.
Desai DV, Dhanani HJ, Kapoor AK, Yeluri SV. HbQ-India in a Sindhi family: an uncommon hemoglobin variant. Lab Hematol. 2004;10(4):212-4.
Panigrahi I, Bajaj J, Chatterjee T, Sakena R, Mahapatra M, Pati HP. Hb Q India: Is it always benign? Am Hematol. 2005;78:245-7.
Desai D, Parmar C, Dhanani HJ, Patel RZ, Master DC. A Rare Case of Co-Existent Hb Q India-Beta Thalassemia Trait. Internet J Hematol. 2006;3(1):1-4.
Lewis MR, Macauley RC, Sheehan PR, Staten MA, Phillips LS, Rasouli N et al. Management of Hemoglobin Variants Detected Incidentally in HbA1c Testing: A Common Problem Currently Lacking a Standard Approach. Diabetes Care. 2017;40(2):8-9.
Bry L, Chen PC, Sacks DB. Effects of hemoglobin variants and chemically modified derivatives on assays for glycohemoglobin. Clin Chem. 2001;47:153-63.
Sukumaran PK, Merchant SM, Desai MP, Wiltshire BG, Lehmann H. Haemoglobin Q India (alpha 64 (E13) aspartic acid histidine) associated with beta-thalassemia observed in three Sindhi families. J medical genetics. 1972;9(4):436.
Wiwanitkit V. Secondary and tertiary structure aberration of alpha globin chain in haemoglobin Q-India disorder. Indian J Pathol Microbiol. 2006;49(4):491-4.
Itano H. A third abnormal hemoglobin associated with hereditary hemolytic anaemia. Proc Nat Acad Sci USA. 1951;37:775-7.
Baglioni C. Abnormal human hemoglobin. VII. Chemicalstudies on hemoglobin D. Biochem Biophys Acta. 1962;59:437-49.
Colaco S, Surve R, Sawant P, Nadkarni A, Ghosh K, Colah R. HbD Punjab/HbQ India Compound Heterozygosity: An Unusual Association. Mediterr J Hematol Infect Dis. 2014;6(1):e2014072.
