Molecular characterization of beta-thalassemia reveals the presence of common mutations in the population of Himalayan region: Garhwal (Uttarakhand), India
Thalassemia is a dreadful heritable hemolytic disease, characterized by a genetic mutation in the hemoglobin subunit beta (HBB) gene. Mutation in HBB gene completely halts the production of the beta-globin protein, which leads to the defective production of functional hemoglobin. The prevalence of this disease is reported only in some specific geographical regions of India. Hence, the aim of this study was to screen the population of Garhwal for beta-thalassemia (β-thalassemia) and thus find out the prevalence in the inhabitants through molecular characterization. For this study, 4,081 individuals were considered, out of which only the ones with elevated HbA2 levels (64) were subjected to molecular characterization. Mutational studies were carried out for the five most common mutations prevalent in the Indian subcontinent, that is, IVS 1-5 G-C, IVS 1-1 G-T, Codon 41/42 (-TCTT), Codon 8/9, and 619 bp deletion. The present study reports a frequency of 0.5% for β-thalassemia mutations among the subjects we have studied. The analysis of mutation spectrum revealed highest prevalence for IVS-1-5 (G-C) (18.75%) followed by Codon 8/9 (12.5%) and IVS-1-1 (G-T) with 6.25%. Codon 41/42 (-TCTT) and 619 bp deletion were found to be absent in our study population.
Agarwal MB & Mehta BC. (1982). Symptomatic beta thalassemia trait (A study of 143 cases. Journal of Postgraduate Medicine, 28(1):4-8.
Agarwal S, Gupta A, Gupta UR, et al. (2003). Prenatal diagnosis in beta-thalassemia: An Indian experience. Fetal Diagnosis and Therapy, 18(5):328-332. https://doi.org/10.1159/000071975
Agarwal S, Pradhan M, Gupta UR, et al. (2000). Geographic and ethnic distribution of beta-thalassemia mutations in Uttar Pradesh, India. Hemoglobin, 24(2):89-97. https://doi.org/10.3109/03630260009003427
Ambekar SS, Phadke MA, Mokashi GD, et al. (2001). Pattern of hemoglobinopathies in Western Maharashtra. Indian Pediatrics, 38(5):530-534.
Baig SM, Azhar A, Hassan H, et al. (2006). Prenatal diagnosis of beta-thalassemia in Southern Punjab, Pakistan. Prenatal Diagnosis, 26(10):903-905. https://doi.org/10.1002/pd.1523
Bajwa H & Basit H. (2022). Thalassemia. In: StatPearls. United States: StatPearls Publishing.
Bandyopadhyay A, Bandyopadhyay S, Basak J, et al. (2004). Profile of beta-thalassemia in eastern India and its prenatal diagnosis. Prenatal Diagnosis, 24(12):992-996. https://doi.org/10.1002/pd.1049
Bashyam MD, Bashyam L, Savithri GR, et al. (2004). Molecular genetic analyses of beta-thalassemia in South India reveals rare mutations in the beta-globin gene. Journal of Human Genetics, 49(8):408-413. https://doi.org/10.1007/s10038-004-0169-9
Bhukhanvala DS, Italia K, Sawant P, et al. (2013). Molecular characterization of β-thalassemia in four communities in South Gujarat--codon 30 (G → A) a predominant mutation in the Kachhiya Patel community. Annals of Hematology, 92(11):1473-1476. https://doi.org/10.1007/s00277-013-1777-2
Black ML, Sinha S, Agarwal S, et al. (2010). A descriptive profile of β-thalassaemia mutations in India, Pakistan and Sri Lanka. Journal of Community Genetics, 1(3):149-157. https://doi.org/10.1007/s12687-010-0026-9
Cao A & Galanello R. (2010). Beta-thalassemia. Genetics in Medicine, 12(2):61-76. https://doi.org/10.1097/GIM.0b013e3181cd68ed
Cao A & Kan YW. (2013). The prevention of thalassemia. Cold Spring Harbor Perspectives in Medicine, 3(2):a011775. https://doi.org/10.1101/cshperspect.a011775
Chakrabarti P, Gupta R, Mishra A, et al. (2005). Spectrum of beta-thalassemia mutations in North Indian states: A beta-thalassemia trait with two mutations in cis. Clinical Biochemistry, 38(6):576-578. https://doi.org/10.1016/j.clinbiochem.2005.03.002
Christopher AF, Kumari A, Chaudhary S, et al. (2013). Unique pattern of mutations in β-thalassemia patients in Western Uttar Pradesh. Indian Journal of Human Genetics, 19(2):207-212. https://doi.org/10.4103/0971-6866.116119
Colah R, Gorakshakar A, Nadkarni A, et al. (2009). Regional heterogeneity of beta-thalassemia mutations in the multi ethnic Indian population. Blood Cells Molecules and Diseases, 42(3):241-246. https://doi.org/10.1016/j.bcmd.2008.12.006
Colah R, Gorakshakar A, Phanasgaonkar S, et al. (2010). Epidemiology of beta-thalassaemia in Western India: Mapping the frequencies and mutations in sub-regions of Maharashtra and Gujarat. British Journal of Haematology, 149(5):739-747. https://doi.org/10.1111/j.1365-2141.2010.08131
De M, Das SK, Bhattacharya DK, et al. (1997). The occurrence of beta-thalassemia mutation and its interaction with hemoglobin E in the Eastern India. International Journal of Hematology, 66(1):31-34. https://doi.org/10.1016/s0925-5710(97)00585-9
De Sanctis V, Kattamis C, Canatan D, et al. (2017). β-Thalassemia Distribution in the old world: An ancient disease seen from a historical standpoint. Mediterranean Journal of Hematology and Infectious Diseases, 9(1):e2017018. https://doi.org/10.4084/MJHID.2017.018
Edison ES, Shaji RV, Devi SG, et al. (2008). Analysis of beta globin mutations in the Indian population: Presence of rare and novel mutations and region-wise heterogeneity. Clinical Genetics, 73(4):331-337. https://doi.org/10.1111/j.1399-0004.2008.00973.x
Garewal G & Das R. (2003). Spectrum of β-thalassemia mutations in Punjabis. International Journal of Human Genetics, 3(4):217-219. https://doi.org/10.1080/09723757.2003.11885855
Grow K, Vashist M, Abrol P, et al. (2014). Beta thalassemia in India: Current status and the challenges ahead. International Journal of Pharmacy and Pharmaceutical Sciences, 6(4):28-33.
Gupta A, Hattori Y, Gupta UR, et al. (2003). Molecular genetic testing of beta-thalassemia patients of Indian origin and a novel 8-bp deletion mutation at codons 36/37/38/39. Genetic Testing, 7(2):163-168. https://doi.org/10.1089/109065703322146894
Haritha P, Lakshmi V, Veerraju P, et al. (2012). Prevalence of haemoglobinopathies among the Konda Kammaras of Vishakhapatnam district, Andhra Pradesh. Journal of Pharmaceutical and Biological Sciences, 2(4):6-8.
Jawahirani A, Mamtani M, Das K, et al. (2007). Prevalence of beta-thalassaemia in subcastes of Indian Sindhis: Results from a two-phase survey. Public Health, 121(3):193-198. https://doi.org/10.1016/j.puhe.2006.10.017
Kazazian HH Jr., Orkin SH, Antonarakis SE, et al. (1984). Molecular characterization of seven beta-thalassemia mutations in Asian Indians. The EMBO Journal, 3(3):593-596. https://doi.org/10.1002/j.1460-2075.1984.tb01853.x
Kukreti R, Dash D, Vineetha KE, et al. (2002). Spectrum of beta-thalassemia mutations and their association with allelic sequence polymorphisms at the beta-globin gene cluster in an Eastern Indian population. American Journal of Hematology, 70(4):269-277. https://doi.org/10.1002/ajh.10117
Kumar R, Arya V, Agarwal S. (2015). Profiling β Thalassemia mutations in consanguinity and nonconsanguinity for prenatal screening and awareness programme. Advances in Hematology, 2015:625721. https://doi.org/10.1155/2015/625721
Little S. (2001). Amplification-refractory mutation system (ARMS) analysis of point mutations. Current Protocols in Human Genetics, Chapter 9:Unit 9.8. https://doi.org/10.1002/0471142905.hg0908s07
Madan N, Sikka M, Sharma S, et al. (1998). Phenotypic expression of hemoglobin A2 in beta-thalassemia trait with iron deficiency. Annals of Hematology, 77(3):93-96. https://doi.org/10.1007/s002770050421
Mohanty D, Colah RB, Gorakshakar AC, et al. (2013). Prevalence of β-thalassemia and other haemoglobinopathies in six cities in India: A multicentre study. Journal of Community Genetics, 4(1):33-42. https://doi.org/10.1007/s12687-012-0114-0
Mulchandani DV, Fulare MB, Zodpey SP, et al. (2008). Prevalence and some epidemiological factors of beta thalassaemia trait in Sindhi community of Nagpur city, India. Indian Journal of Public Health, 52(1):11-15.
Nagar R, Sinha S & Raman R. (2015). Haemoglobinopathies in eastern Indian states: A demographic evaluation. Journal of Community Genetics, 6(1):1-8. https://doi.org/10.1007/s12687-014-0195-z
Old JM, Khan SN, Verma I, et al. (2001). A multi-center study in order to further define the molecular basis of beta-thalassemia in Thailand, Pakistan, Sri Lanka, Mauritius, Syria, and India, and to develop a simple molecular diagnostic strategy by amplification refractory mutation system-polymerase chain reaction. Hemoglobin, 25(4):397-407. https://doi.org/10.1081/hem-100107877
Ou Z, Li Q, Liu W, et al. (2011). Elevated hemoglobin A2 as a marker for β-thalassemia trait in pregnant women. The Tohoku Journal of Experimental Medicine, 223(3):223-226. https://doi.org/10.1620/tjem.223.223
Panigrahi I & Marwaha RK. (2007). Mutational spectrum of thalassemias in India. Indian Journal of Human Genetics, 13(1):36-37. doi:10.4103/0971-6866.32034
Piplani S, Manan R, Lalit M, et al. (2013). NESTROFT-A valuable, cost effective screening test for beta thalassemia trait in North Indian Punjabi population. Journal of Clinical and Diagnostic Research, 7(12):2784-2687. https://doi.org/10.7860/JCDR/2013/6834.3759
Sahoo SS, Biswal S, & Dixit M. (2014). Distinctive mutation spectrum of the HBB gene in an urban Eastern Indian population. Hemoglobin, 38(1):33-38. https://doi.org/10.3109/03630269.2013.837394
Satpute SB, Bankar MP, & Momin AA. (2012). The prevalence of β-thalassemia mutations in South Western Maharashtra. Indian Journal of Clinical Biochemistry, 27(4):389-393. https://doi.org/10.1007/s12291-012-0230-y
Shah PS, Shah ND, Ray HS, et al. (2017). Mutation analysis of β-thalassemia in East-Western Indian population: A recent molecular approach. The Application of Clinical Genetics, 10:27-35. https://doi.org/10.2147/TACG.S127531
Thomas S, Srivastava A, Jeyaseelan L, et al. (1996). NESTROFT as a screening test for the detection of thalassaemia and common haemoglobinopathies-An evaluation against a high performance liquid chromatographic method. Indian Journal of Medical Research, 104:194-197.
Usman M, Moinuddin M, GhaniR, et al. (2009). Screening of five common beta thalassemia mutations in the Pakistani population: A basis for prenatal diagnosis. Sultan Qaboos University Medical Journal, 9(3):305-310.
Vaz FE, Thakur CB, Banerjee MK, et al. (2000). Distribution of beta-thalassemia mutations in the Indian population referred to a diagnostic center. Hemoglobin, 24(3):181-194. https://doi.org/10.3109/03630260008997526
Verma IC, Saxena R, Thomas E, et al. (1997). Regional distribution of beta-thalassemia mutations in India. Human Genetics, 100(1):109-113. https://doi.org/10.1007/s004390050475
Weatherall D & Clegg J. (2001b). The Thalassaemia Syndromes. Hoboken: Blackwell Science Ltd., https://doi.org/10.1002/9780470696705
Weatherall DJ & Clegg JB. (2001a). Inherited haemoglobin disorders: An increasing global health problem. Bulletin of the World Health Organization, 79(8):704-712.
World Health Organization, UNICEF, UNU. Iron deficiency Anaemia: Assessment, Prevention and Control, a Guide for Programme Managers. Geneva, World Health Organization, 2001. Available from: https://www.who.int/publications/m/ item/iron-children-6to23--archived-iron-deficiency-anaemia-assessment-prevention-and-control [Last accessed on 2022 Nov 30].
World Health Organization. (2008). Worldwide Prevalence of Anaemia 1993-2005: WHO Global Database on Anaemia. Geneva: World Health Organization. Available from: https:// www.apps.who.int/iris/handle/10665/43894 [Last accessed on 2022 Nov 30].