Deteksi Mutasi Langka, Delesi 619 bp, pada Gen Beta-Globin dari Etnis Melayu Mahasiswa Fakultas Kedokteran Universitas YARSI

Kenconoviyati Kenconoviyati, Kinasih Prayuni, RW Susilowati, Rika Yuliwulandari, Abdul Salam M. Sofro

Abstract


Beta-thalassemia merupakan gangguan hematologis autosomal yang secara genetis mengakibatkan berkurangnya sintesis beta-globin di hemoglobin. Beta-talasemia sebagian besar disebabkan oleh mutasi titik, insersi atau delesi dalam gen beta-globin yang terletak pada lengan pendek kromosom 11. Organisasi Kesehatan Dunia (WHO) memperkirakan terdapat sekitar 1,5% dari populasi global (80-90 juta orang) adalah pembawa ?-thalassemia. Tidak ada studi komprehensif untuk mendeteksi pembawa beta-thalassemia di Indonesia, terutama untuk mutasi delesi 619 bp, yang mencakup ekson 3 dan memiliki prevalensi yang tinggi. Kami menggunakan metode gap-PCR yang di-kombinasikan dengan metode elektroforesis gel untuk memper-kirakan adanya mutasi delesi 619 bp pada 48 siswa Fakultas Kedokteran Universitas YARSI dengan etnis Melayu. Analisis Blast hasil sekuensing dari ketiga sampel menunjukkan bahwa terdapat similaritas 98% antara hasil amplifikasi dengan ke daerah gen beta-globin pada kromosom 11 (No. Aksesi U01317.1). Berdasarkan hasil visualisasi elektroforesis gel, semua produk PCR dari 48 sampel, menunjukkan bahwa semua sampel tidak membawa mutasi delesi 619 bp yang ditunjukkan dengan ukuran produk PCR yang sama dari semua sampel, yaitu berukuran 1.457 bp dan 2.291 bp dari PCR I dan 1.212 bp dari PCR II.

Beta-thalassaemia is an autosomal haematological disorder resulting in a genetically deficient synthesis of the ?-globin chain in haemoglobin. It is mostly caused by point mutations, a small deletions or insertions within the beta-globin gene which is located as a cluster on the short arm of chromosome 11. The World Health Organization has estimated that about 1.5% of the global population (80 to 90 million people) were carriers of ?-thalassemia. There are no comprehensive study to detect carrier of ?-thalassemia in Indonesia especially for 619 bp deletion mutation, which encompasses exon 3, that has greater prevalence. We used gap-PCR combined with gel electrophoresis methods to roughly screen the presence of major indel mutation in 48 Medical Faculty, Universitas YARSI students with Malay ethnic. To validate whether the PCR product obtained is the beta-globin gene, a direct sequencing of 3 PCR products were performed. The Blast analysis of the sequence was also done using NCBI database. The result showed that the PCR products obtained in this study showed 98% identity to human beta-globin gene region on chromosome 11 (No. Acc. U01317.1). In the electrophoresis of all PCR products of 48 samples, the result showed that all the samples did not carry any major indel mutation showing by the presence of similar length of PCR products in gel electrophoresis, which has 1.457 bp and 2.291 bp product from PCR I and 1.212 bp product from PCR II.

 


Keywords


Beta-thalassemia; gen Beta-globin; Etnis Melayu; gap-PCR; elektroforesis gel; Delesi 619 bp

Full Text:

Download File

References


Baysal E, Sharma S, Wong SC, Jogessar VB, & Huisman TH 1994. Distribution of beta-thalassemia mutations in three Asian Indian populations with distant geographical locations. Hemoglobin, 18:201-209.

Chan OTM, Westover KD, Dietz L, Zehnder JL and Schrijver I 2010. Comprehensive and Efficient HBB Mutation Analysis for Detection of ? Hemoglobinopathies in a Pan Ethnic Population. America Journal Clinical Pathology, 133: 700-707.

Fucharoen S & Winichagoon P 2007. Prevention and control of thalassemia in Asia. Asian Biomedicine, 1(1):1–6.

Galanello R and Origa R 2010. Beta-thalassemia. Orphanet Journal of Rare Disease, 21: 11.

Hassan S, Ahmad R, Zakaria Z, Zulkafli Z and Abdullah WZ 2013. Detection of ?-globin Gene Mutations Among ?-thalassaemia Carriers and Patients in Malaysia: Application of Multiplex Amplification Refractory Mutation System–Polymerase Chain Reaction Malays Journal Medical Science, 20(1): 13-20.

Lin M, Jiao JW, Zhan XH, Zhan XF, Pan MC, Wang JL et al. 2014. High Resolution Melting Analysis: A Rapid Screening and Typing Tool for Common b-Thalassemia Mutation in Chinese Population. Plos One, 9: 1-7.

Munshi A, Anandraj MP, Joseph J, Shafi G, Anila AN & Jyothy A 2009. Inherited hemoglobin disorders in Andhra Pradesh, India: a population study. Clin Chim Acta, 400:117-119.

Orkin SH, Old JM, Weatherall DJ, & Nathan DG 1979. Partial deletion of beta-globin gene DNA in certain patients with beta 0-thalassemia. Proc Natl Acad Sci USA, 76:2400-2404.

Orkin SH, Kolodner R, Michelson A, & Husson R 1980. Cloning and direct examination of a structurally abnormal human beta 0-thalassemia globin gene. Proc Natl Acad Sci USA, 77:3558-3562.

Rund D, & Rachmilewitz E 2005. Beta-thalassemia. The New England Journal of Medicine, 15: 1135-1146.

Sheth JJ, Sheth FJ, Pandya P, Priya R, Dayla S, Thakur C et al. 2008. Beta-thalassemia mutations in western India. Indian J Pediatr, 75:567-570.

Shih HC, Er TK, Chang TJ, Chang TS, Li TC, & Chang JG 2009. Rapid identification of HBB gene mutation by high-resolution melting analysis. Clinical Biochemistry 42: 1667–1676.

Thein SL, Old JM, Wainscoat JS, Petrou M, Modell B, & Weatherall DJ 1984. Population and genetic studies suggest a single origin for the Indian deletion beta thalassaemia. Br J Haematol, 57:271-278.

Thein SL 2005. Genetic modifiers of ?-thalassemia. Haematologica, 90(5): 649-660.

Wang W, Kham SK, Yeo GH, Quah TC, & Chong SS 2003. Multiplex minisequencing screen for common Southeast Asian and Indian beta-thalassemia mutations. Clinical Chemistry, 49:209-218.




DOI: https://doi.org/10.33476/jky.v23i2.98

Refbacks

  • There are currently no refbacks.


Copyright (c) 2015 Journal of Medicine Faculty

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

__________________________________________________________________________________________

Copyright of YARSI Medical Journal.

Powered by OJS.

Creative Commons License

This work is licensed under a Creative Commons Attribution- NonCommercial 4.0 International License.