Laboratory evaluation of hemolytic anemia

Author of a master‘s thesis. Dalia Vertelytė The title of the master‘s thesis. Laboratory evaluation of hemolytic anemia. The aim: To evaluate changes of laboratory tests during hemolytic anemia and relate them with of type of hemolytic anemia. Objectives: To evaluate hematology test results, lactate dehydrogenase (LDH) activity, haptoglobin (HAPTO) concentration and direct antiglobulin test (DAT) in patients with hemolytic status; To compare groups of patients according to levels of LDH activity to LDH upper reference limit ratio LDH/URL; To evuluate the changes of laboratory parameters in patients with hemolytic anemia according to presence of clone of paroxysmal norturnal hemoglobinuria (PNH). Patients and methods. Patients with disorders of hemopoiesis were included in the study, then patients tested for HAPTO and LDH were chosen, and additional selection was done – 80 patients with HAPTO concentration lower than 360 mg/l were left for the final analysis. 26 patients were included in flow cytometry analysis. Hematology tests were done using automated hematologic s XE–5000 (Sysmex, Japan). The biochemistry tests were run using clinical system – SYNCHRON UniCel® DxC 800 (Beckman Coulter, USA) and DAT were tested by column hemagglutination technique (Biorad, USA). The immunophenotyping of RBC and WBC was carried out for identification of PNH clone using flow cytometre BD FacsCanto (BD Immunocytometry Systems, USA). Results. The prevalence of negative DAT results didn‘t differ between male and female patients with hemolytic anemias. RBC, HGB, HCT values were significantly lower and TBIL – significantly higher in patients with LDH/URL1.5. Spearman‘s correlative test revealed reverse significant correlation between LDH and HAPTO (R=-0,239; p=0,033). The anemia markers –RBC and HGB significantly negatively correlated with the LDH. Frequency of markers characteristic to PNH (combination of DAT negative results and increased LDH) didn‘t differ between male and female patients. Patients having PNH clone had 12-fold increased of LDH/URL and lower HAPTO, RBC, HGB compared to patients without PNH clone. Conclusions. Patients with hemolytic statuses had decreased erythrocyte and hemoglobin values and increased lactate dehydrogenase activity. There were no differences in laboratory values between men and women. Patients with LDH/URL1.5 had significantly reduced erythrocyte, hemoglobin, haptoglobin and increased total bilirubin (due to increase of indirect bilirubin). LDH activity was directly related to anemia expression. These findings didn‘t depend on gender. Changes of markers indicating anemia and hemolysis were more pronounced in patients with hemolysis and PNH clone. There were no changes and differences of inflammatory markers – neutrophils, as well neutrophil to lymphocyte ratio – between investigated groups