Identification of spurious hemolysis in anticoagulated blood with Sysmex XE-2100 and Siemens Advia 2120.

Background: Various degrees of hemolysis might occur during collection, processing and storage of blood bags or blood tubes for hematological testing. In both circumstances, the identification of hemolysis is challenging since the centrifugation process is not required. The aim of this study was to identify simple hematological parameters that would help identify the presence of hemolysis in anticoagulated blood. Methods: Twenty tubes containing K(2)EDTA anticoagulated blood were randomly selected from outpatient samples and divided in two aliquots. The former was immediately analyzed, whereas the latter was subjected to mechanical hemolysis by aspirating whole blood two times through a very fine needle to generate mechanical hemolysis. Both aliquots were tested on Advia 2120 and Sysmex XE-2100. Results: The double aspiration of the blood through the fine needle caused a remarkable hemolysis with significant decrease of red blood cell (RBC) count (-17 +/- 11%; p<0.01), hematocrit (-18 +/- 12%; p<0.01) and reficulocytes (-24 +/- 13%; p<0.01), but not of hemoglobin, white blood cell or platelet counts. A remarkable increase of immature platelet fraction (IPF) on XE-2100 and RBC ghosts on Advia 2120 was observed in the hemolyzed samples, whereas RBC fragments did not vary significantly. A significant correlation was also observed between hemolysis and reticulocyte count (r = 0.823; p<0.01), IPF (r = -0.502, p = 0.024) and RBC ghosts (r = -0.711; p<0.01). Receiver Operating Characteristic (ROC) curve analysis demonstrated a good performance of both IPF and RBC ghosts (i.e., AUCs 0.91 and 0.96, respectively) for distinguishing non-hemolyzed from hemolyzed specimens. Conclusions: The absolute values of both IPF and RBC ghosts perform efficiently for distinguishing hemolyzed from non-hemolyzed specimens, although neither reached 100% sensitivity and specificity. Nevertheless, the demonstration that both parameters significantly increase after hemolysis can be reliably used to distinguish hemolyzed from non-hemolyzed blood. (Clin. Lab. 2012;58:801-804. DOI: 10.7754/Clin.Lab.2011.111001)