Application of the polymerase chain reaction to the diagnosis of sickle cell disease in Iran

Background – Sickle cell anemia is one of the most common heritable hematologic diseases affecting humans. Detection of the single base pair mutations at codon 6 of the beta-globin gene is important for the prenatal diagnosis of sickle cell anemia and sickle cell disease. We applied the polymerase chain reaction technology to detect sickle cell patients and heterozygous carriers in a group of patients suspected of sickle cell disease.Methods – The sample was composed of 45 normal individuals and 45 unrelated sickle cell disease out-patients from Hematology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. All patients were interviewed. Results of their medical histories, physical examination, and the hematologic analysis were recorded. The blood samples were collected in EDTA and genomic DNA was extracted from leucocytes. An amplified 110 base pair fragment of beta-globin gene containing codon 6 was digested with the restriction enzyme MS II, and electrophoresed in 3% agarose. Results – We have established the technical condition for detection of sickle cell disease using a PCR assay. Thirteen patients having hemoglobin SS (Hb SS) and 32 patients in the heterozygous state (Hb AS) were identified. We confirmed that the normal controls had the Hb AA genotype. Conclusion – This amplification method is rapid, sensitive, and simple, and has the application which is important for the prenatal diagnosis of sickle cell disease.