Familial erythrocytosis is a hereditary disease characterized by an increased mass of erythrocytes and increased levels of hemoglobin and hematocrit. Due to hyperviscosity of the blood, erythrocytosis can cause health problems such as thrombotic complications. Bisphosphoglycerate mutase (BPGM) is a gene associated with familial erythrocytosis type 8 (ECYT8). The BPGM gene encodes a transcript for bisphosphoglycerate mutase (BPGM), which is a protein that catalyzes the conversion of 1,3-bisphosphoglycerate (1,3-BPG) to 2,3-bisphosphoglycerate (2,3-BPG), the allosteric effector of hemoglobin. Mutations in the BPGM gene can cause altered structure and decreased function of BPGM protein and therefore lower concentrations of 2,3-BPG in the blood, causing an increased affinity of hemoglobin for oxygen. Hence, the oxygen delivery to the tissues decreases, causing chronic tissue hypoxia and consequent ECYT8. The master's thesis was performed in cooperation with the Specialized Hematology Laboratory of the Clinical Department of Hematology of the University Medical Center in Ljubljana. Using bioinformatical analysis, we first identified regions for genetic analysis. Then, we successfully established a genetic test to verify the presence of variants in the regions of the first and second exons of the BPGM gene, by PCR optimization and Sanger sequencing. The established genetic test was then confirmed in two Slovenian patients with suspected familial erythrocytosis. According to the analysis of the acquired nucleotide sequences, the patients had no variations in the tested regions of the BPGM gene, thus excluding the possibility of ECYT8. In the future, the established genetic test will be used in combination with other tests, for routine analysis of the BPGM gene in the diagnosis of familial erythrocytosis in Slovenia.
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