Rapid and simultaneous detection of β- thalassemia point mutations by reverse hybridization strip assay among Egyptian patients

Authors

  • Osama Shalaan Molecular Diagnostics and Therapeutics Department, Genetic Engineering and Biotechnology Research institute (GEBRI), University of Sadat City, Egypt.
  • Moustafa Sakr Molecular Diagnostics and Therapeutics Department, Genetic Engineering and Biotechnology Research institute (GEBRI), University of Sadat City, Egypt.
  • Ahmed Daif Molecular Diagnostics and Therapeutics Department, Genetic Engineering and Biotechnology Research institute (GEBRI), University of Sadat City, Egypt
  • Khalil Elhalfawy Molecular Diagnostics and Therapeutics Department, Genetic Engineering and Biotechnology Research institute (GEBRI), University of Sadat City, Egypt

DOI:

https://doi.org/10.54987/jobimb.v4i1.282

Keywords:

β-Thalassemia, mutations, β-Globin Strip Assay (reverse dot-blot PCR), Egyptian

Abstract

Thalassemia is a wide range hereditary disease with high incidence in Egypt along with the high frequency of consanguineous marriages. Investigation the heterogeneity, molecular basis, and natural history are the most effective methods to deal with the thalassemia to develop effective method for management and prevention including the prenatal diagnosis. The current study aims to detect the most common β -globin gene mutations in Egypt among β-thalassemic patients by using PCR based reverse hybridization method (StripAssay) for the most prevalent 22 β-globin gene mutations in the Mediterranean population in an attempt to estimate the incidence of each mutation, and an attempt to improve our control strategy of β- thalassemia. This study included a total of 37 confirmed β- thalassemia ethnic Egyptian patients (23 males and 14 females) out of them 17 patients were a thalassemia major and 20 were a thalassemia intermediate. Evaluation of β-thalassemia mutations revealed that, the presence of 9 different β-globin mutations. The most frequent mutation was IVS 1-110[34%], IVS 1-6(23.5%), IVS 1-1(19%), Codon 27[6.5%], IVS 2-848[6.5%], IVS 2-745[2.1%] and IVS 2.1 [2.5%], Codon 39[4%]), and IVS 1.5 [1.5%]. IVS 1-110[G>A] is the commonest homozygous mutation found in 6 out of 14 homozygous cases accounting for about 43% of homozygous mutations while, IVS 1-110[G>A]/ IVS 1-6[T>C] is the commonest heterozygous mutation found in 6 out of 23 heterozygous cases accounting for about 26% of heterozygous mutations. In conclusion, β-Globin Strip Assay is useful rapid screening method for β-globin gene mutations which can provide an insight into the detection of individual patients, to take the proper measurement, and thus could be used as a basis for genetic counseling and prenatal diagnosis.

References

Bain BJ. The alpha, beta, delta and gamma thalassaemias and related conditions. In Haemoglobinopathy Diagnosis, 2nd Edition. Wiley-Blackwell. Oxford; 2006:63-138.

Weatherall DJ, Clegg JB, Higgs DR, Wood WG. The hemoglobinopathies. In:Scriver CR, Beaudet Al, Sly WS, Valle D, Vogelstein B, editors. The metabolic and molecular bases of inherited disease (OMMBID). Chapter 101. New York, NY: McGraw-Hill, 2002. Available at: www.ommbid.com. AccessedDecember 31, 2007

Al-Allawi NA, Jubrael JM, Hughson M. Molecular characterization of beta-thalassemia in the Dohuk region of Iraq. Hemoglobin 2006; 30:479–486.

El-Beshlawy A, Kaddah N, Ragab L, Hussein I, Mouktar G, Moustafa A, El -Raouf E, Hassaballa N, Gaafarand T, El-Sendiony H. Thalassemic prevalence and status in Egypt. Proceedings of the annual meeting of the American Pediatric Society, San Francisco, CA, USA, (1-4 may 1999); abstract 102.

El-Hashemite N, Petrou M, Khalifa AS, et al. Identification of novel Asian Indian and Japanese mutations causing β-thalassemia in the Egyptian population. Hum Genet. 1997;99(2):271-274.

Ahmed S, Saleem M, Modell B, Petrou M. Screening extended families for genetic hemoglobin disorders in Pakistan. N Engl J Med. 2002;347:1162-1168.

Ahmed S. Prenatal diagnosis of β-thalassemia: 12 years’ experience at a single laboratory in Pakistan. Prenatal Diag. 2007;27:1224-1227.

Naseem S, Ahmed S, Vahidy F. Impediments to prenatal diagnosis of beta thalassemia: experiences from Pakistan. Prenatal Diag. 2008;28:1116-1118.

Omar A, Abdel Karim E, El Gendy W, Marzouk I, Wagdy M. Molecular basis of beta thalassemia in Egypt. Egypt J Immunol. 2005;12(1):15–24.

Colah R, Gorakshakar A, Nadkarni A, Phanasgaonkar S, Surve R, Sawant P, Mohanty D, Ghosh K. Regional heterogeneity of ß-thalassemia mutations in the multi ethnic Indian population. Blood Cells Mol Dis. 2009;42(3):241-246.

Edison ES, Shaji RV, Devi SG, Moses A, Viswabandhya A, Mathews V, George B, Srivastava A, Chandy M. Analysis of ß globin mutations in the Indian population: presence of rare and novel mutations and region-wise heterogeneity. Clin Genet. 2008;73(4):331-337.

Black ML, Sinha S, Agarwal S, Colah R, Das R, Bellgard M, Bittles AH. A descriptive profile of β-thalassemia mutations in India, Pakistan and Sri Lanka. J Commun Genet. 2010;1:149-157.

Başak AN. The molecular pathology of ß-thalassemia in Turkey: The Boğazıçı University experience. Hemoglobin. 2007;31:233-241.

Akhavan-Niaki H, Derakhshandeh-Peykar P, Banihashemi A, Mostafazadeh A, Asghari B, Ahmadifard MR, Azizi M, Youssefi A, Elmi MM. A comprehensive molecular characterization of beta thalassemia in a highly heterogeneous population. Blood Cells Mol Dis. 2011;47:29-32.

Girodon E, Ghanem N, Vidaud M, Riou J, Martin J, Galacteros F, Goossens M. Rapid molecular characterization of mutations leading to unstable hemoglobin beta-chain variants. Ann Hematol. 1992;65:188–192.

Zhang YH, McCabe LL, Wilborn M, Therrell BL Jr, McCabe ER. Application of molecular genetics in public health: improved follow-up in a neonatal hemoglobinopathy screening program. Biochem Med Metab Biol. 1994; 27–35.

Herrmann MG, Dobrowolski SF, Wittwer CT. Rapid beta-globin genotyping by multiplexing probe melting temperature and color. Clin Chem. 2000;46:425–428.

Hussein G, Fawzy M, Serafi TE, Ismail EF, Metwally DE, Saber MA, Giansily M, Schved JF, Pissard S, Martinez PA. Rapid detection of ï¢-thalassemia alleles in Egypt using naturally or amplified created restriction sites and direct sequencing: a step in disease control. Hemoglobin 2007;31(1):49.

El-Beshlawy A and Youssry I. Prevention of hemoglobinopathies in Egypt. Hemoglobin 2009; 33 (1):14-20.

EL-Fadaly, A. Abd-Elhameed, E. Abd-Elbar, M. El-Shanshory. Accuracy of Reverse Dot-Blot PCR in Detection of Different b-Globin Gene Mutations. Indian J Hematol Blood Transfus 2015.

Barrai I, Vullo C. Assessment of prospective genetic counseling in the Ferrara area. Am J Med Genet. 1980;6(3):195–204.

Soliman HH, Kabbash IA, El-Shanshory MR, Nagy HM, Abdou SH. Evaluation of immune status against hepatitis B in children with thalassemia major in Egypt. J Microbiol Infect Dis. 2012;2:44–49.

Galanello R, Barella S, Satta S, Maccioni L, Pintor C, Cao A. Homozygosity for nondeletion delta-beta(0) thalassemia resulting in a silent clinical phenotype. Blood 2002;100:1913–1914.

El mezayen A. Kotb S, Ahmed M, Sadek NA, Abdalla EM. β-globin mutations in Egyptian patients with β-thalassemia. Lab Med. 2015;46:8-13.

El-Beshlawy, A. El-Shekha, M. Momtaz, F. Said, M. Hamdy, O. Osman, S. Meshaal, T. Gafaar and M. Petrou. Prenatal diagnosis for thalassaemia in Egypt: what changed parents’ attitude. Prenat Diagn. 2012; 32,1–6.

Jiffri E, Bogari N, Zidan K, Teama S, Elhawary NA. Molecular updating of β-thalassemia mutations in the upper Egyptian population. Hemoglobin, 2010;34(6):538–547.

El-Gawhary S, El-Shafie S, Niazi M, Aziz M, El-Beshlawy A. Study of β-thalassemia mutations using the polymerase chain reaction amplification refractory mutation system and direct DNA sequencing techniques in a group of Egyptian thalassemia patients. Hemoglobin, 2007;31: 63-69.

Guardiola J, Rojo V, Ramos G. Influence of particle size, fluidization velocity and relative humidity on fluidized bed electrostatics. J Electrost. 1996;37(1-2):1–20.

Murtomaa M, Savolainen M, Christiansen L, Rantanen J, Laine E, Yliruusi J. Static electrification of powders during spray drying. J Electrost. 2004;62(1):63–72.

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Published

31.07.2016

How to Cite

Shalaan, O., Sakr, M., Daif, A., & Elhalfawy, K. (2016). Rapid and simultaneous detection of β- thalassemia point mutations by reverse hybridization strip assay among Egyptian patients. Journal of Biochemistry, Microbiology and Biotechnology, 4(1), 11–14. https://doi.org/10.54987/jobimb.v4i1.282

Issue

Vol. 4 No. 1 (2016)

Section

Articles

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