Molecular Genetic Confirmatory Testing for the Sickle Cell Anaemia using Restriction Fragment Length Polymorphism (RFLP) in Sudan
Journal Title: Journal of Medical Science And clinical Research - Year 2015, Vol 3, Issue 2
Abstract
Background: Molecular diagnosis of the disease, genetic and family studies of patients becomes an important tool for management of sickle cell disease patients.. In this study we compare the efficiency of agarose gel electrophoresis techniques with recently developed molecular biology methods. (RFLP) for the diagnosis of in Sudanese patients with SCD. Methods and Results: This study was conducted in Khartoum state between 2007 to 2008.in hundred and fifty patients with homozygous SCD of age between 6 month –40 years. Questionnaires were used to collect demographic and clinical data. About 3 ml of venous ant coagulated blood were collected for DNA extraction. Each extracted DNA samples was subjected to PCR reaction. .Polymerase chain reaction was carried out for DNA amplification. , The results of RFLP In this method after the amplification of DNA with primers1 (mutant), primer 2 (normal primer), the 281bp fragment was achieved, after that we digested this fragment with DdeI restriction enzyme in two fragment (200bp&81 bp) so +/+ shows that our sample is normal and we had 200 & 81 bp both of them but we couldn't see 80 bp so we just had 200 bp! For heterozygous carrier we had 3 bands 200&281&81 that we saw just 2 bands (200 and 281), that was -/+! for homozygous disease we had just one band 281 bp that shows our restriction sits disturb and our enzyme couldn't digest it The study thus demonstrates the accuracy and precision of Restriction Fragment Length Polymorphism (RFLP)techniques in molecular genetic confirmatory testing for the sickle cell anaemia
Authors and Affiliations
Nadia, M . M
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