Nguyen Thu Trang, Le Thi Thu Ha, Nguyen Thuy Ngan, Trieu Tien Sang, Pham Anh Thuy Duong, Vo Thi Thuong Lan

Main Article Content


Abstract: Beta (β)-thalassemia is the most common genetic disease of anemia caused by mutations on beta globin gene. In Vietnam, there is a high frequency of β -thalassemia carriers with a prevalence ranging from 1.5 to 25.0 % in the different ethnic groups. The most common mutations are the nonsense in codon (CD) 17 (A>T), codon 26 (G>A) and the frameshift at codons 41/42 (-TTCT). The polymerase chain reaction-amplification refractory mutation system (PCR-ARMS) is challenged by using a great number of primer sets for detecting normal and mutated alleles. Reverse dot-blot hybridization using oligonucleotide probes can simultaneously detect allelic specific mutations on a membrane. In this study, we designed oligonucleotide probes specific to the three most common mutations in CD17, CD26 and CD41/42, and used them to optimize hybridization conditions at 68 0C in 6xSSC hybridization buffer, 2XSSC washing buffer for detecting homozygous or hetezygous alleles of beta globin gene in fifteen individuals of 5 families. Our results were consistent with those detected by PCR-AMRS method, indicating that oligonucleotide probes created by this study was specific, and that the reverse dot-blot hybridization using these oligo probes was convenient for analysis of beta thalassemia disease inVietnam.

Keywords: Beta globin gene, beta thalassemia disease,  polymerase chain reaction-amplification refractory mutation system (PCR-ARMS), reverse dot-blot  analysis.


[1] S. Fucharoen, P. Winichagoon, Haemoglobinopathies in Southeast Asia, Indian J Med 134 (2011) 498.
[2] M.A. Abdulla, I. Ahmed, A. Assawamakin, J. Bhak, S.K. Brahmachari, G.C. Calacal, et al., Mapping human genetic diversity in Asia, Science 3262 (2009) 1541.
[3] A. Cao, R. Galanello., Beta-thalassemia, Genet Med 12 (2010) 61.
[4] A.A. Settin, M.M. Al-Haggar, M. Neamatallah, A.M. Al-Said, M.M. Hafez, Detection of beta-thalassemia mutations using primer-specific amplification compared to reversed dot blot hybridization technique in Egyptian cases,. Haema 9 (2006) 401.
[5] B. Quoc Tuyen, N. Kim Hoa, N. Thi Loi, N. Dinh Luong, B. Bich, N. Thi Khiem, Study of hemoglobin diseases in the hematological department of Bach Mai hospital 1963–1973, Hematol Transfus 17 (1974) 135.
[6] B.K. Hoa, N.Q. Cuong, Screening of mutations causing beta thalassemia in Northern region of Vietnam, Tạp chí Nghiên cứu Y học 40 (2006) 53.
[7] L.T.T. Ha, N.T.P. Mai, N.T.M. Hương, N.D. Ngọc, D.B.Trực, N.T. Liêm, N.T.T. Sinh, Application of ARMS-PCR for prenatal diagnosis of beta thalassemia at Vietnam National Hospital of Pediatrics, http://www.
[8] D. Filon, A. Oppenheim, E.A. Rachmilewitz, R. Kot, D. Ba Truc, Molecular analysis of beta thalassemia in Vietnam, Hemoglobin 24 (2000) 99.
[9] P. Winichagoon, V. Saechan, R. Sripanich, C. Nopparatana, S. Kanokpongsakdi, A.Maggio, S. Fucharoen, Prenatal diagnosis of beta-thalassemia by reverse dot-blot hybridization, Prenat Diagn 19 (1999) 428.
[10] D. Li, C. Liao, J. Li, Y. Huang, X. Xie , J. Wei, S. Wu, Prenatal diagnosis of beta-thalassemia by reverse dot-blot hybridization in southern China, Hemoglobin 30 (2006) 365.
M. Lin, T-Y. Zhong, Y-G. Chen, J-Z. Wang, J-R.Wu et al., Molecular epidemiological characterization and health burden of thalassemia in Jiangxi province, P. R. China, PLoS ONE 9(7): e101505. 2014, doi:10.1371/ journal. pone. 0101505.