Background: In Africa, the Sickle Cell Disease, SCD, is the most common inherited hematological disease with a high mortality rate at age one to five years. This disease was discovered early in Sudan. The peak occurrence of SCD is among the population from Western Sudan. The objective of this study is to demonstrate the accuracy and precision of Amplification Refractory Mutation System - Polymerase Chain Reaction,)ARMS- PCR ( technique for the diagnosis of SCD in Sudanese patients .
Methods and results: This study was conducted in Khartoum State between 2005 to 2008.in hundred and fifty patients with homozygous SCD from 6 months up to 40 years of age. Questionnaires were used to collect demographic and clinical data. About 3 ml of venous anti coagulated blood were collected for DNA extraction. Each extracted DNA samples was subjected to Allele-specific-PCR. Allele-specific-PCR primers were designed to amplify the HbS allele and the normal HbA allele when paired with a common reverse primer.Separate PCR reactions were set up for the HbA and HbS primer pairs, and each reaction contained control primers. Polymerase chain reaction was carried out for DNA amplification. Visible bands were seen at 207 bp regions for the AA, AS, SS genotypes yielding 207bp. Results from agarosegel electrophoresis also showed the same ARMS PCR results. Moreover,ARMS-PCRis potential method to permit rapid analysis of any known mutation in genomic DNA, it allows genotyping solely by inspection of reaction mixtures after agarose gel electrophoresis
The study thus demonstrates the accuracy and precision of Amplification Refractory Mutation System - polymerase chain reaction)ARMS- PCR) techniques in diagnosis of sickle cell anaemia.
Keywords: Amplification Refractory Mutation System, polymerase chain reaction, sickle cell anaemia, electrophoresis.