![]() ![]() Pramoonjago P, Harahap A, Taufani RA, Setianingsih I, Marzuki S, Harahap A (1999) Rapid screening for the most common beta thalassaemia mutations in south east Asia by PCR based restriction fragment length polymorphism analysis (PCR-RFLP). Nicosia, Cyprusīaig SM (2007) Molecular diagnosis of beta-thalassemia by multiplex ARMS-PCR: a cost effective method for developing countries like Pakistan. Old J, Harteveld CL, Traeger-Synodinos J, Petrou M, Angastiniotis M, Galanello R (2012) Prevention of thalassaemias and other haemoglobin disorders: volume 2: laboratory protocols, 2nd edn. Yavarian M, Harteveld CL, Batelaan D, Bernini LF, Giordano PC (2001) Molecular spectrum of beta-thalassemia in the Iranian Province of Hormozgan. Najmabadi H, Karimi-Nejad R, Sahebjam S, Pourfarzad F, Teimourian S, Sahebjam F et al (2001) The beta-thalassemia mutation spectrum in the Iranian population. Īkhavan-Niaki H, Derakhshandeh-Peykar P, Banihashemi A, Mostafazadeh A, Asghari B, Ahmadifard MR et al (2011) A comprehensive molecular characterization of beta thalassemia in a highly heterogeneous population. Ībolghasemi H, Amid A, Zeinali S, Radfar MH, Eshghi P, Rahiminejad MS et al (2007) Thalassemia in Iran: epidemiology, prevention, and management. Our designed RSB test is a rapid, sensitive and cost-effective method for screening of regional specific beta-thalassemia mutations in the Khuzestan population of Iran, which might be extended for the detection of any desired pathogenic changes.Ĭao A, Galanello R (2010) Beta-thalassemia. Hybridization of each PCR products on the nylon membrane with probe pairs revealed specific bands with expected signal intensity without any background. In the next step, DNA samples from homozygous affecting individuals were subjected for multiple PCR. The optimal probe concentration for each mutation varied from 25 to 50 pmol. For the best probe concentration, we made a serial dilution of probe pairs for each mutation. Finally, we developed the membrane by chemically colorimetric reaction using nitro-blue tetrazolium-5-Bromo-4-chloro-3-indolyl phosphate. The PCR products were hybridized to immobilized oligonucleotide probes on the membrane at the appropriate temperature. In the next step, a multiplex-polymerase chain reaction (PCR) performed for the amplification of the entire HBB gene using labelled 5′-biotinylated primers. We designed normal and mutant oligonucleotide probes for each selected mutation and fixed them on positively charged nylon membrane. Here, we developed reverse slot blot (RSB) assay for the simultaneous detection of six common pathogenic changes in the HBB gene (-88, -28, IVSII-745, IVSII-848, Codon 6 for HbC, Codon 6 for HbS) in the Khuzestan Province of Iran. Therefore, it is essential, depending on the ethnicity and local frequency of changes, to develop a rapid and accurate method for molecular diagnosis of beta-thalassemia. The screening of beta-thalassemia minor and major individuals and prenatal diagnosis is important for familial planning. Beta-thalassemia is the most frequent hemoglobin disorder in Iran resulting from disrupting mutations in the beta globin ( HBB) gene that causes decreased or complete absent of beta-globin chains. ![]()
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