Abstract
SummaryBackgroundThe aim of this study is to establish the contribution of blood cells subtypes on hemolysis.MethodsSeparated blood cell subtype suspensions prepared with blood from 10 volunteers were serially diluted to obtain different concentrations of cell suspensions. The cells were fully lysed and cell hemolysates were added (1:20) to aliquots of serum pool. Thus, seven serum pools with different concentrations of interferent were obtained for each blood cell subtype. Biochemical parameters and serum indices were measured by an autoanalyzer. As cell lysis markers, free hemoglobin was measured by spectrophotometry while myeloperoxidase and ᵝ-thromboglobulin were measured by enzyme immunoassay. The percent changes in analyte levels of the serum pools were evaulated by Wilcoxon Signed Rank Test and compared with clinical thresholds defined for each test.ResultsThe clinical thresholds were exceeded in lactate dehydrogenase, potassium, aspartate aminotransferase, creatine kinase, magnesium, total protein, total cholesterol, inorganic phosphate, glucose for red blood cells (RBC); lactate dehydrogenase, aspartate aminotransferase, total protein, inorganic phosphate and glucose for platelets (PLT). Free hemoglobin was significantly correlated with RBC (r=0.999; p=0.001), while myeloperoxidase and b thromboglobulin showed no significant correlation to white blood cells (WBC) and PLT, respectively.ConclusionsThe effect of RBC hemolysis in serum on the routine biochemical tests are clearly established, yet, additional studies are required in order to verify this kind of effects of PLT and WBC hemolysis.
Highlights
Hemolyzed samples make up 3% of the routine samples sent to clinical laboratories and are responsible for 39–69% of the unsuitable specimens, this is 5 times more than the second most frequent reason [1]
Free hemoglobin was measured by spectrophotometry while myeloperoxidase and b-thromboglobulin were measured by enzyme immunoassay
The percent changes in analyte levels of the serum pools were evaulated by Wilcoxon Signed Rank Test and compared with clinical thresholds defined for each test
Summary
Hemolyzed samples make up 3% of the routine samples sent to clinical laboratories and are responsible for 39–69% of the unsuitable specimens, this is 5 times more than the second most frequent reason [1]. Hemolysis may occur in vivo, the in vitro hemolysis in the preanalytical phase is the major problem faced by clinical laboratories [2, 3]. Elucidation of the interference mechanisms of hemolysis is necessary for more precise solutions of the problem of in vitro hemolysis. Hemolyzed samples interfere with the test results by several mechanisms such as compositional interference due to the difference between the intra- and extracellular concentration of the analytes, signal interference in instrumental measurements and chemical interference in analytical reactions. These interference mechanisms might coexist in varying combinations [4]. The rejection of the test results of hemolyzed samples may lead to a delay in diagnosis threatening the patient’s safety, and the request of an additional sample increases the workload and the cost (6 –10)
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