Variant Hemoglobin Spectrum By Cation Exchange High Performance Liquid Chromatography: A Study 0f 2035 Subjects
Keywords:
Haemoglobinopathies, Thalassemia, Cation exchange high performance liquid chromatography, ScreeningAbstract
Introduction
Hemoglobinopathies and thalassemia are hereditary disorders of hemoglobin (Hb) affecting mankind at prevalent regional level. Automated cation exchange high performance liquid chromatography is being increasingly used as the initial diagnostic method for identifying normal and abnormal hemoglobin variants.
Methodology
Total 2035 sample received and studied. All samples run on cation exchange high performance liquid chromatography machine by BIO-RAD.
Results:
Total 386(18.96%) cases had abnormal hemoglobin fractions. Beta thalassemia major cases were 17(0.83%). Beta thalassemia intermedia cases were 4(0.19%). Sickle cell trait (heterozygous Hb S) cases were 96(4.71%). Double heterozygous for sickle cell-beta thalassemia cases were 14(0.68%). There were 06 cases (0.29%) of Hb D heterozygous. There were one each case of Hb E heterozygous and Hb E-beta thalassemia (0.04% each). Hb F was raised in 7 adult cases (0.34%).
Conclusion:
To conclude, cation exchange high performance liquid chromatography is less time consuming, cost effective, labor saving, reproducible, accurate, sensitive and specific method to detect hemoglobinopathies and thalassemia. Most of the abnormal cases are diagnosed with this method, with few inconclusive cases require further genetic and molecular workup.
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DOI:Â 10.21276/APALM.1393
References
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24. Bencaiova G, Burkhardt T, Krafft A, Zimmermann R. Screening for β-thalassaemia trait in anaemic pregnant women. Gynecologic and obstetric investigation. 2006 Mar 1;62(1):20-7.
25. Gupta AD. Abrogation of macrocytosis in pernicious anemia by β-thalassemia does not mask the diagnosis of vitamin B12 deficiency. American journal of hematology. 2002 Sep 1;71(1):61-2.
26. Sachdev R, Dam AR, Tyagi G. Detection of Hb variants and hemoglobinopathies in Indian population using HPLC: report of 2600 cases. Indian journal of pathology and microbiology. 2010 Jan 1;53(1):57.
27. Rao S, Kar R, Gupta SK, Chopra A, Saxena R. Spectrum of haemoglobinopathies diagnosed by cation exchange-HPLC & modulating effects of nutritional deficiency anaemias from north India. Indian Journal of Medical Research. 2010 Nov 1;132(5):513.
28. Dolai TK, Dutta S, Bhattacharyya M, Ghosh MK. Prevalence of hemoglobinopathies in rural Bengal, India. Hemoglobin. 2012 Feb 1;36(1):57-63.
29. Mukhopadhyay D, Saha K, Sengupta M, Mitra S, Datta C, Mitra PK. Spectrum of hemoglobinopathies in West Bengal, India: a CE-HPLC Study on 10407 subjects. Indian Journal of Hematology and Blood Transfusion. 2015 Mar 1;31(1):98-103.
30. Jain BB, Roy RN, Ghosh S, Ghosh T, Banerjee U, Bhattacharya SK. Screening for thalassemia and other hemoglobinopathies in a tertiary care hospital of West Bengal: implications for population screening. Indian journal of public health. 2012 Oct 1;56(4):297.
2. Kutlar F. Diagnostic approach to hemoglobinopathies. Hemoglobin. 2007 Jan 1;31(2):243-50.
3. WHO executive board EB118/5, 118th Session Report by the Secretariat on Thalassaemia and other haemoglobinopathies: Prevalence of Haemoglobinopathies. 11 May 2006:1-8.
4. Varawalla NY, Old JM, Sarkar R, Venkatesan R, Weatherall DJ. The spectrum of βâ€thalassaemia mutations on the Indian subcontinent: the basis for prenatal diagnosis. British journal of haematology. 1991 Jun 1;78(2):242-7.
5. Balgir RS. The genetic burden of hemoglobinopathies with special reference to community health in India and the challenges ahead. Indian J Hematol Blood Transfus. 2002;20(1):2-7.
6. Gupta PK, Kumar H, Kumar S, Jaiprakash M. Cation exchange high performance liquid chromatography for diagnosis of haemoglobinopathies. Medical Journal Armed Forces India. 2009 Jan 31;65(1):33-7.
7. Wild BJ, Stephens AD. The use of automated HPLC to detect and quantitate haemoglobins. International Journal of Laboratory Hematology. 1997 Sep 1;19(3):171-6.
8. Higgins TN, Ridley B. Tentative identification of hemoglobin variants in the Bio-Rad VARIANT II Hb A 1C method. Clinical biochemistry. 2005 Mar 31;38(3):272-7.
9. Phelan L, Bain BJ, Roper D, Jury C, Bain K. An analysis of relative costs and potential benefits of different policies for antenatal screening for beta thalassaemia trait and variant haemoglobins. Journal of clinical pathology. 1999;52(9):697-700.
10. Riou J, Godart C, Hurtrel D, Mathis M, Bimet C, Bardakdjian-Michau J, Préhu C, Wajcman H, Galactéros F. Cation-exchange HPLC evaluated for presumptive identification of hemoglobin variants. Clinical chemistry. 1997;43(1):34-9.
11. Bio–Rad VARIANT II, beta thalassemia short program. Instruction Manual. 2003:10.
12. Working Party of the General Haematology Task Force of the British Committee for Standards in Haemotology. Guideline: The laboratory diagnosis of haemoglobinopathies. British Journal of Haematology. 1998;101:783-92.
13. Chakrabarti I, Sinha SK, Ghosh N, Goswami BK. Beta-Thalassemia Carrier Detection by NESTROFT: An Answer in Rural Scenario?. Iranian Journal of Pathology. 2012 Jan 1;7(1):19-26.
14. Gorakshakar AC, Colah RB. Is RBC discrimination index suitable for differentiating between α-and β-thalassemias?. Indian journal of human genetics. 2011 Sep 1;17(3):115.
15. Dangi CB, Sajid M, Sawke GK, Ambhore J. Sickle cell hemoglobinopathies in district Bhopal. Indian journal of human genetics. 2010 May 1;16(2):100.
16. Yousafzai YM, Khan S, Raziq F. Beta-thalassaemia trait: Haematological parameters. J Ayub Med Coll Abbottabad. 2010;22:84-6.
17. Joutovsky A, Hadzi-Nesic J, Nardi MA. HPLC retention time as a diagnostic tool for hemoglobin variants and hemoglobinopathies: a study of 60000 samples in a clinical diagnostic laboratory. Clinical chemistry. 2004 Oct 1;50(10):1736-47.
18. Somervaille T. Disorders of Hemoglobin: Genetics, Pathophysiology, and Clinical Management. Journal of the Royal Society of Medicine. 2001; 94(11):602-603.
19. Dash S, Huisman TH. Hb [A. sub. 2] in subjects with Hb D. Clinical chemistry. 1998 Nov 1;44(11):2381-3.
20. Bain BJ. Haemoglobinopathy diagnosis. 2nd ed. Oxford: Blackwell Publishing Ltd; 2006.
21. Ou CN, Rognerud CL. Diagnosis of hemoglobinopathies: electrophoresis vs. HPLC. Clinica chimica acta. 2001 Nov 30;313(1):187-94.
22. El-Agouza I, Abu Shahla A, Sirdah M. The effect of iron deficiency anaemia on the levels of haemoglobin subtypes: possible consequences for clinical diagnosis. Clinical & laboratory haematology. 2002 Oct 1;24(5):285-9.
23. Madan N, Sikka M, Sharma S, Rusia U. Haematological parameters and HbA2 levels in beta-thalassaemia trait with coincident iron deficiency. Indian journal of pathology & microbiology. 1998 Jul;41(3):309-13.
24. Bencaiova G, Burkhardt T, Krafft A, Zimmermann R. Screening for β-thalassaemia trait in anaemic pregnant women. Gynecologic and obstetric investigation. 2006 Mar 1;62(1):20-7.
25. Gupta AD. Abrogation of macrocytosis in pernicious anemia by β-thalassemia does not mask the diagnosis of vitamin B12 deficiency. American journal of hematology. 2002 Sep 1;71(1):61-2.
26. Sachdev R, Dam AR, Tyagi G. Detection of Hb variants and hemoglobinopathies in Indian population using HPLC: report of 2600 cases. Indian journal of pathology and microbiology. 2010 Jan 1;53(1):57.
27. Rao S, Kar R, Gupta SK, Chopra A, Saxena R. Spectrum of haemoglobinopathies diagnosed by cation exchange-HPLC & modulating effects of nutritional deficiency anaemias from north India. Indian Journal of Medical Research. 2010 Nov 1;132(5):513.
28. Dolai TK, Dutta S, Bhattacharyya M, Ghosh MK. Prevalence of hemoglobinopathies in rural Bengal, India. Hemoglobin. 2012 Feb 1;36(1):57-63.
29. Mukhopadhyay D, Saha K, Sengupta M, Mitra S, Datta C, Mitra PK. Spectrum of hemoglobinopathies in West Bengal, India: a CE-HPLC Study on 10407 subjects. Indian Journal of Hematology and Blood Transfusion. 2015 Mar 1;31(1):98-103.
30. Jain BB, Roy RN, Ghosh S, Ghosh T, Banerjee U, Bhattacharya SK. Screening for thalassemia and other hemoglobinopathies in a tertiary care hospital of West Bengal: implications for population screening. Indian journal of public health. 2012 Oct 1;56(4):297.
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Sarvaiya AN, Chauhan SC. Variant Hemoglobin Spectrum By Cation Exchange High Performance Liquid Chromatography: A Study 0f 2035 Subjects. Ann of Pathol and Lab Med [Internet]. 2017 Jul. 5 [cited 2024 Dec. 26];4(3):A297-302. Available from: https://pacificejournals.com/journal/index.php/apalm/article/view/apalm1393
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