Background: The aim of this study is to establish the contribution of blood cells subtypes on hemolysis.
Methods: Separated 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.
Results: The 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.
Conclusions: The 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.
Uvod: Cilj ovog proučavanja je da se utvrdi uticaj podtipova krvnih ćelija na hemolizu.
Metode: Suspenzija izdvojenih podtipova ćelija krvi pripremljena od krvi 10 dobrovoljaca serijski je razblažena kako bi se dobile razlićite koncentracije ćelijskih suspenzija. ]elije su upotpunosti lizirane i ćelijski hemolizati su dodavani (1:20) alikvotima serumskog pula. Na ovaj način dobijeno je sedam serumskih pulova sa različitim koncentracijama interferenata. Biohemijski parametri mereni su na autoanalizatoru. Slobodni hemoglobin, kao marker hemolize, meren je spektrofotometrijski, dok su mijeloperoksidaza i β-tromboglobulin mereni enzimskim imunoodređivanjem. Procenat promene nivoa analita u serumskim pulovima procenjivan je Wilcoxon Signed Rank Testom i upoređivan sa klinički definisanim za svaki test.
Rezultati: Klinički značajne promene nađene su za laktat dehidrogenazu, kalijum, aspartat aminotransferazu, kreatin kinazu, magnezijum, ukupne proteine, ukupni holesterol, neorganski fosfat, i glukozu za eritrocite (RBC); laktat dehidro genazu, aspartat aminotransferazu, ukupne proteine, neorganski fosfat i glukozu za trombocite (PLT). Slobodni hemoglobin je bio u znaćajnoj korelaciji sa RBC (r = 0,999; p = 0,001), dok mijeloperoksidaza i β-tromboglobulin nisu bili u značajnoj korelaciji sa leukocitima (WBC) i trombocitima (PLT).
Zaključak: Utvrđen je znaćajan efekat RBC hemolize u serumu na rutinske biohemijske testove, dok su još potrebna dodatna ispitivanja za potvrdu ovakvih efekata hemolize PLT i WBC.
Keywords: biochemical test; free hemoglobin; hemolysis; interference; myeloperoxidase.