[Application of the polymerase chain reaction to the diagnosis of sickle cell anemia in Venezuela].

Application of polymerase chain reaction technology to detect sickle cell patients and heterozygous carriers in a group of patients suspect for sickle cell disease was carried out. The sample was composed of 102 normal individuals, and 102 unrelated out patients who were attending in the Hematology Service at the Maracay Central Hospital in the State of Aragua in Venezuela. All patients were interviewed. Results of their medical histories and the physical examination, made during the clinical visit, were recorded. The blood samples were collected in EDTA by venopuncture and genomic DNA was extracted from leucocytes. An amplified 536 base pairs fragment of the beta-globin gene containing codon 6, was digested with an isoschizomer of Mst II, Bsu36 I and electrophoresed in 3% agarose. We have established the technical conditions in our laboratory for the detection of sickle cell disease using a PCR assay. 32 patients having haemoglobin SS (HbSS) and 70 patients in the heterozygous state (HbAS) were identified. We confirm that the normal controls have the HbAA genotype. The standardization of a highly sensitive and specific diagnostic test for sickle cell disease permited us to identify the normal controls, the homozygotes and heterozygotes. This methodology is one of the fundamental technical bases for establishing a newborn screening programme in the Central Region of Venezuela and also has application in research related with other genetic diseases that affect the Venezuelan people.