*1 Department of Medical Research (Pyin Oo Lwin Branch) 2 Department of Medical Research 3 University of Medicine (Mandalay) 4 Myanmar Academy of Medical Science
Beta thalassemia is one of the most common genetic disorders worldwide with a defect in haemoglobin synthesis including Myanmar. This study was conducted to determine the proportion of high school students who have heterozygous ?-thalassaemia including Hb E and different types of common beta globin gene mutations by multiplex polymerase chain reaction. This community-based cross-sectional descriptive study was done in Myanmar adolescents in 2017. A total 290 apparently healthy high school students in which male 140 (48.3%) and female 150 (51.7%) studying in Basic Education High School, Anisakhan, Pyin Oo Lwin Township, Mandalay District, Myanmar were involved. Thalassaemia screening tests were done by combined NESTROFT/DCIP precipitation and confirmed test was done by Isoelectric Focusing (IEF). PCR- based genetic confirmatory by using mutant genes IVS 1-1, CD 41/42, IVS 1-5, CD 17, and nt-28. Total 131 cases (131/290, 45.2%) were positive with either of the two screening tests. These cases were considered as thalassaemia trait (carrier) and/or haemoglobin E carrier. Isoelectric focusing was done to find Hb E by which 45 cases (45/131, 34.3%) were found to have Hb E carrier. Remaining 86 cases were further run multiplex- PCR to detect beta thalassaemia carrier. Total 44 out of 86 cases were ?-thalassaemia traits (25 CD 41/42 (-TCTT), 13 IVS 1-5 (G-C), 1 IVS 1-1 (G-T); 5 compound heterozygous (IVS 1-5(G-C) /CD 41/42 (-TCTT)). CD 17 (A-T) and nt 28 (A-G) mutations were not detected in this study. NESTROFT screening test has 87% sensitivity and 82% sensitivity for the detection of Hb E carrier and ?-thalassaemia carrier respectively. DCIP screening test has 67.1% sensitivity and 68% sensitivity for the Hb E carrier and ?-thalassaemia carriers respectively. Combined both screening tests have 100% sensitivity for the detection of Hb E and ?-thalassaemia carriers. This study has highlighted that both screening tests should be used for population screening of thalassaemia and haemoglobinopathies in Myanmar.
Khin Moe Aung,Aye Aye Lwin,Haung Naw,Nan Cho Nwe Mon,Hnin Ohmar Soe ,Myitzu Tin Oung,Nilar Wynn, Ne Win."Detection of Common Beta Globin Gene Mutations on Heterozygous Beta Thalassemia Including Haemoglobin E among High School Students". International Journal of Computer Engineering In Research Trends (IJCERT) ,ISSN:2349-7084 ,Vol.7, Issue 01,pp.1-7,January - 2020, URL :https://ijcert.org/ems/ijcert_papers/V7I101.pdf,
: beta thalassaemia, beta globin gene, haemoglobin E, high school students
1. Origa, R, ‘Beta Thalassaemia, In: Pagon RA, Adam MP, Ardinger HH, et al., editors. Gene Reviews® [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2016, Retrieved 2016 August 18. Retrieved 2016August10, 2014. http://www.ncbi.nlm.nih.gov/books/NBK1426/
2. Galanello, R & Origa, R, ‘Beta-thalassaemia’, Orphanet Journal of Rare Diseases, vol. 5, pp. 1-15, 2010.
3. TIF Guidelines (Thalassaemia International Federation Guidelines), ‘Guideline for the Management of Transfusion Dependent Thalassaemia’, 3rd ed, pp. 12-17, 2012.
4. Origa, R, Galanello, R, Ganz, T, Giagu, N, Maccioni, L, Faa, G & Nemeth, E, ‘Liver Iron Concentrations and Urinary Hepcidin in Beta-Thalassaemia’, Haematologica, vol. 92, pp. 583–588, 2007.
5. Winichagoon, P, Thitivichianlert, A, Lebnak, T, Piankijagum, A & Fucharoen, S, , ‘Screening for the Carriers of Thalassemias and Abnormal Hemoglobins at the Community Level Southeast Asian’ , J Trop Med Public Health, vol. 33, pp. 145-150, 2002.
6. Fucharoen, G, Sanchaisuraiya, K, Sae-ung, N, Dangwibul, S & Fucharoen, S, ‘A Simplified screening Stragey for Thalassaemias and Haemoglobin E in Rual Communities in Southeast Asia’, Bull World Health Organ, vol. 82, pp. 364-372, 2004.
7. Old, J, Harteveld, CL, Traeger-Synodinos, J, Petron, M, Angastiniotis, M & Galanello, R, ‘Prevention of Thalassaemias and Other Haemoglobin Disorders’: Volume 2: Laboratory Protocols [Internet]. 2nd ed. Nicosia, Cyprus: Thalassaemia International Federation;2012.Retrieved2016August9, 2012 https://www.ncbi.nlm.nih.gov/books/NBK190576/
8. Aung-Than-Batu, Khin-Kyi-Nyunt & Hla-Pe, ‘The Thalassaemia in Burma’, Union of Burma, J life Sciences, vol. 1, pp. 241-247, 1968.
9. Harano, K, Ne-Win & Harano, T, ‘Molecular Aspects of Transfusion Dependent Thalassaemic Children in Myanmar: Analysis of Common - Thalassaemia in Myanmar by Amplification Refractory Mutation System (ARMS) ’, Kawasaki Med J, vol. 26, pp. 161-164, 2000.
10. Yi-Yi-Tin, ‘Phenotypic and Genotypic Heterogeneity of Beta-Thalassaemia Intermedia in Myanmar’, Ph. D (Pathology) Thesis, University of Medicine (2), Yangon, Republic of the Union of Myanmar, Retrieved 2016 August 18, from University of Medicine, Mandalay Digital Thesis, 2004.
11. Sein-Win, ‘Clinical Spectrum and Genetic Heterogeneity of E-? Thalassaemia Intermedia in Myanmar Adults at Yangon General Hospital’, Ph.D. (Clincal Hematology) Thesis; University of Medicine (I), Yangon, Republic of the Union of Myanmar, Retrieved 2016 August 18, from University of Medicine, Mandalay Digital Thesis, 2010.
12. Brown, JM, Thein, SL, Weatherall, DJ & Khin-Mar-Mar, ‘The Spectrum of ?- Thalassaemia in Burma’, Br J Haematol, vol. 81, pp. 574-578, 1992.
13. Ne-Win, Harano, T., Harano, K., Thein-Thein-Myint, Rai-Mra, Okada, S., Shimono K. & Aye-Aye-Myint, ‘A Wider Molecular Spectrum of ?-Thalassaemia in Myanmar’, Br J Haematol, vol. 117, pp. 988-992, 2002.
14. Ne Win.. ‘Clinical and Molecular Heterogeneity of Hemoglobine E-Beta Thalassemia and Beta-Thalassemia Major in Myanmar’, Thesis for the degree of Doctor of Philosophy (PhD) Pathology, University of Medicine I, Yangon, Republic of the Union of Myanmar, Retrieved 2016 August 12, from University of Medicine, Mandalay Digital Thesis, 2002.
15. Hassan, S, Ahmad, R, Zakaria, Z, Zulkafli, Z & Abdullah, WZ, ‘Detection of ?-Globin Gene Mutations Among ?-thalassaemia Carriers and Patients in Malaysia: Application of Multiplex Amplification Refractory Mutation System–Polymerase Chain Reaction’, Malays J Med Sci., vol. 20, pp. 13-20, 2013.
16. Boonyawat, B., Monsereenusorn, C. & Traivaree, C. Molecular Analysis of Beta Globin Gene Mutations among Thai Beta-Thalassaemia Children: Results from a Single Center Study. The Application of Clinical Genetics. 7, 253–258, 2014.
17. Tongprasert, F., Sirichotiyakul, S., Piyamongkol, W., Tongsong, T. Sesitivity and Specificity of Erythrocyte Osmotic Fragility Test for Screening of ?-Thalassaemia-1 and ?-Thalassaemia in Pregnant Women. Gynaecol Obstet Invest, 69, 217- 20, 2010.
18. Viroj, W. Osmotic Fragility Test for Screening for Thalassaemia in Thai Pregnant Subjects: A Re-evaluation. J Haema. 7(2), 205-7, 2004.
19. Mamtani, M., Das, K. & Jawahirani A. Is NESTROFT sufficient for mass screening for ?-thalassemia trait? J Med Screen. 14, 169–73, 2007.
20. Singh, SP. & Gupta, SC. Effectiveness of red cell osmotic fragility test with varying degrees of saline concentration in detecting ?-thalassaemia trait. Singapore Med J. 49(10), 823-6, 2008.
21. Yazdani, SM., Ahmed, S. & Abbas F. One Tube Osmotic Fragility Test: A Screening Test for Microcytic Red Cells. Pak Armed Forces Med J. 58(1), 45-50, 2008.
22. Chakrabarti, I., Sinha, SK., Ghosh, N. & Goswami, BK. Beta-Thalassemia Carrier Detection by NESTROFT: An Answer in Rural Scenario? Iranian Journal of Pathology. 7 (1), 19 – 26, 2012.
23. Bobhate, S., Gaikwad, S. & Bhaledrao, T. NESTROFT as a Screening Test for Detection of ?-Thalassemia Trait. Indian J Pathol Microbiol. 45 (3), 265-7, 2002.
24. Sangkitporn, S., Sangnoi, A., Supangwiput, O., Tanphaichitr, VS. Validation of Osmotic Fragility and Dichlorophenol Indophenol Precipitation Test for Screening of Thalassaemia and Hb E. South East Asian J Trop Med Public Health. 36(3), 1538-42, 2005.
25. Jopang Y., Thinkhamrop, B., Puangpruk R. & Netnee, P. False positive rates of thalassaemia screening in rural clinical setting: 10- year experience in Thailand. South East Asian J Trop Med Public Health. 40(3), 576-580, 2009.
26. Sanchaisuriya, K., Fucharoen, S., Fucharoen G., et al. A Reliable Screening Protocol for Thalassemia and Hemoglobinopathies in Pregnancy: an Alternative Approach to Electronic Blood Cell Counting. Am J Clin Pathol. 123, 113-8, 2005.
27. Weatherall, DJ. & Clegg, JB. The Thalassaemia Syndromes’, Oxford: Blackwell Scientific Publications, 4th Ed, 770-80, 2001.
28. Insiripong, S., Yingsitsiri, W., Boondumrongsagul, J. & Noiwatanakul, J. Prevalence of Thalassaemia Traits in People without Anaemia or Microcytosis. J haematol Transfus Med. 24, 25-9, 2014.
29. Ambekar, SS., Phadke, MA., Balpande, DN., Mokashi, GD., Khedkar, VA., Bankar, MP., Gambhir, PS., Bulakh, PM. & Basutkar, DG. The Prevalence and Heterogeneity of Beta Thalassemia Mutations in the Western Maharashtra Population: A Hospital Based Study. International Journal of Human Genetic.1 (3), 219-223, 2001.
30. Huisman, THJ., Carver, MFH. & Baysl, E. A Syllabus of Thalassa Mutations. The Augusta, GA: Sickle Cell Anaemia Foundation. (http://globin.cse.psu.edu)., 1997.