Daria Pašalić
Department of Medical Chemistry, Biochemistry and Clinical Chemistry
Zagreb University School of Medicine
Šalata ul 2.
10 000 Zagreb, Croatia
Phone +385 (1) 4590 205; +385 (1) 4566 940
E-mail: dariapasalic [at] gmail [dot] com

Useful links

P03-1 (Oral presentation)

Paradinović K, Milić M, Majetić-Cetina N. PO3-1: Case report: complete blood count in severe lipemic sample. Biochemia Medica 2009;19(Suppl 1):S114.
Department for clinical laboratory diagnostics, Osijek University Hospital, Osijek, Croatia
Corresponding author:paradinovic [dot] ksenija [at] kbo [dot] hr
Background: Severe lipemia is a known interference that can falsely elevate a hemoglobin result obtained from colorimetric hemoglobin method. When hemoglobin is falsely increased, the MCH and MCHC are also falsely increased.
Aim: Determination and correction of hemoglobin concentration and calculated parameters in severe lipemic sample.
Material and methods: Complete blood count was determined on ADVIA 2120 which determinates hemoglobin by two methods: colorimetric and directly measuring CHCM to calculate cellular hemoglobin. Since the results between these two methods were significant, we determined plasma hemoglobin and triglyceride in serum.
Results: Colorimetric hemoglobin concentration was 158.0 g/L. Cellular hemoglobin was 93.0 g/L. Triglycerides in serum were 120.66 mmol/L. Hemoglobin in plasma was 89.0 g/L which we used in correcting hemoglobin in whole blood. After correction hemoglobin concentration was 92.3 g/L.
Conclusion: Using CHCM to calculate hemoglobin is unaffected by lipemia.
P03-2 (Oral presentation)
Šimek S, Županić D, Mujagić R, Honović L. PO3-2: The percentage of hypocromic red blood cells as a marker of functional iron deficiency in hemodialysis patients. Biochemia Medica 2009;19(Suppl 1):S115.
Laboratory of Medical Biochemistry, Pula General Hospital, Pula, Croatia
Corresponding author:danijela [dot] zupanic [at] pu [dot] t-com [dot] hr
Introduction: Hypocromic red blood cells are sensitive marker of sideropenic erythropoiesis and functional anemia in hemodialysis patients treated with erythropoietin. The aim of this study was to show the relationship between the results of some hematological and biochemical parameters obtained from hemodialysis patients on their period checkup.
Materials and methods: The study included 44 patients. Laboratory diagnostics included determination of hemoglobin (Hb), percentage of hypocromic red blood cells (%HYPO), reticulocytes, iron, TIBC, UIBC and ferritin. Hematological parameters were determined on cell counter ADVIA 120 (Bayer, USA), iron, TIBC and UIBC on Olympus AU400 analyzer (Olympus, Japan) and ferritin on ARCHITECT i2000 SR analyzer (Abbott, USA).
Results: In accordance with the guidelines for treating anemia in patients with chronic kidney failure in examinated group, Hb < 110 g/L was found in 29 (66%) patients, %HYPO > 2.5 in 13 (29%), %HYPO > 10 in 2 (4.5%), ferritin < 100 ng/mL in 14 (31.8%) patients. For the assessment of diagnostic significance of %HYPO in discovering the functional iron deficiency, ROC analysis is used with cut off ferritin concentration < 100 ng/mL as a criteria. Calculated area under the curve is 0.72 (95% CI = 0.565-0.845) and the corresponding optimal cut off value of %HYPO > 2% indicate a possible iron deficiency with a diagnostic sensitivity and specificity 64.2%, and 76.7%.
According to Spearman correlation a significant relationship between %HYPO and Hb (P = 0.046) and %HYPO and UIBC (P < 0.001) was found.
Conclusion: Measured parameters show a significant connection between %HYPO and Hb and %HYPO and UIBC. However, according to accepted guidelines for treatment of anemia the recommended target values for hemoglobin and ferritin have not been reached.
Bernt Živković T, Kralik Oguić S. PO3-3: Tromboelastometry in pregnant women with preeclampsia. Biochemia Medica 2009;19(Suppl 1):S116.
Clinical Institute for Laboratory Diagnostics, Clinical Zagreb Hospital Centre, Zagreb, Croatia
Corresponding author:tajanaberntzivkovic [at] net [dot] hr
Introduction: Pregnancy is considered a hypercoagulable state. There is much interest in the relationship between coagulation status and complications of pregnancy. Increased incidence of thromboembolic phenomena have been reported in pregnant women. Pre-eclampsia and eclampsia may complicate some pregnancies and is often associated with abnormalities of hemostasis. The tromboelastograph (TEG) has been proposed as a useful, inexpensive tool to screen for patients with hypercoagulable state. A TEG measures whole blood coagulation and fibrinolysis.
Aim: The purpose of this study was to document tromboelastografic (TEG) changes in preeclampsia or eclampsia and to compare these resuts with the same results in health, normal, term pregnant women.
Patients and methods: A TEG was performed in 28 normal, term pregnant women (35-41 weeks of pregnancy) and in 34 preeclamptic women admitted to the delivery (35-39 weeks of pregnancy) using native whool blood (Rotem delta, Pentapharm GmbH). The TEG variables included CT - clotting time, CFT - Clot formation time, MCF - Maximum Clot Firmness, alpha angle and percentage of reduction in MCF at 60 minutes.
Results: TEG parameters for both groups showed that term pregnant women as well as preeclamptic women were in hypercoagulable state, but no significant differences between these two groups were observed.
Conclusion: Although standard laboratory tests are still necessary to detect coagulation abnormalities the TEG is useful as it is compact, easily located within the delivery suite, relatively easy to use and produces initial results very quickly. Preeclampsia is associated with endothelial damage, impaired production of prostacycline and increased deposition of fibrin within the vascular bed. The TEG does not measure these local changes, but can be used to determine the effects on overall clotting.
Coen Herak D, Miloš M, Zadro R. PO3-4: Sensitivity of the Platelet Function Analyzer (PFA-100) as a screening test for laboratory diagnosis of von Willebrand disease. BiochemiaMedica 2009;19(Suppl 1):S117.
Clinical Institute of Laboratory Diagnosis, Zagreb University School of Medicine and Clinical Hospital Center, Zagreb, Croatia
Corresponding author:desireecoen [at] yahoo [dot] com
Introduction: Many studies have shown high sensitivity of Platelet Function Analyzer 100 (PFA-100) to von Willebrand disease (VWD).
Methods: We evaluated sensitivity of PFA-100 to different degrees of VWF:RCo/VWF:Ag deficiency, by measuring closure times (CTs) with collagen/epinephrine (CEPI) and collagen/ADP (CADP) in groups with different VWF:RCo/VWF:Ag values: group with normal values (> 50%), group 1 (40-50%), group 2 (30-39.9%), group 3 (20-29.9%) and group 4 (< 20%).
Results: In the group with normal VWF:RCo values (N = 145), normal CTs were obtained in 82.1% cases with CEPI, 86.2% with CADP, and 74.5% cases with both tests. In the group with normal VWF:Ag values (N = 141), normal CTs were obtained in 74.5% cases with CEPI, 78.0% with CADP and 66.7% cases with both tests. In the groups with decreased VWF:RCo values, the calculated sensitivities for CEPI-CT, CADP-CT and both tests, were as follows: group 1 (N = 15): 60%, 46.7% and 40%, respectively; group 2 (N = 10): 50%, 60% and 50%, respectively; group 3 (N = 13): 76.9%, 92.3% and 76.9%, respectively; group 4 (N = 18): 100% for CEPI-CT, CADP-CT and both. In groups with decreased VWF:Ag values, the calculated sensitivities for CEPI-CT, CADP-CT and both tests, were as follows: group 1 (N = 8): 62.5%, 87.5% and 62.5%, respectively; group 2 (N = 7): 85.7%, 100% and 85.7%, respectively; group 3 (N = 4) and 4 (N = 8): 100 for CEPI-CT, CADP-CT and both.
Conclusion: PFA-100 as a screening test for VWD shows higher sensitivity towards lower VWF:RCo/VWF:Ag values, with 100% sensitivity for patients with values below 20%.
Pauković Sekulić B, Bošnjak N, Sapunar A, Tandara L, Salamunić I. PO3-5: Comparison of decreased platelet counts on different haematology analyzers. BiochemiaMedica 2009;19(Suppl 1):S118.
Department of Laboratory Diagnostics, Split University Hospital Centre, Split, Croatia
Corresponding author:branka [dot] paukovic-sekulic [at] st [dot] htnet [dot] hr
Introduction: Although haematology analyzers provide the high precision and accuracy in determining the number of platelets, they cannot completely distinguish the platelets from other cell fragments. The aim of the study was to examine the correlation of platelet counts and own empiric cognitions about the comparability of results from different analyzers.
Materials and methods: The platelet counts were determined parallel by optical method on analyzers Advia 120 (Bayer, USA), Sysmex XE-2100 (Toa, Kobe, Japan), Cell-Dyn Sapphire (Abbott Diagnostics, USA) and by impedance method on analyzer Sysmex XE-2100. In the study were tested 30 normal subjects, 50 patients with the platelets 21-100 x 109/L and 20 patients with the platelets below 20 x 109/L toward values obtained on Advia 120. In normal subjects and patients with platelets 21-100 x 109/L the correlation coefficient (r) and coefficient of determination (R2) between analyzers were r = 0.931; R2 = 0.866. In patients with platelets below 20 x 109/L coefficients between analyzers were: Advia-Sysmeximp r = 0.781; R2 = 0.611; Advia-Sysmexopt r = 0.616; R2 = 0.380; Advia-Sapphireopt r = 0.756; R2 = 0.572. Passing-Bablok regression were satisfied for the normal subjects but it were not satisfied for the patients with platelets 21-100 x 109/L between: Advia-Sysmexopt (95% CI, b = 0.850 to 0.986); Advia-Sapphireopt (95% CI, a = -10.833 to -2.134) and in patients with platelets below 20 x 109/L between Advia-Sapphireopt (95% CI, a = -8.167 to -0.875). 95 % limits of agreements of the mean differences obtained from the summarised Bland-Altman diagram were for platelets below 20 x 109/L between: Advia-Sysmeximp -8.829 to 4.528; Advia Sysmexopt -8.341 to 9.441; Advia- Sapphireopt -10.998 to 1.598.
Results: The study showed the clinical significant disagreement of results between analyzers in patients with decreased platelet counts. In the daily work, it is recommend observe these patients on the same analyzer, or making the additional verification of result by microscope or flow cytometry.
Coen Herak D, Miloš M, Zadro R.PO3-6: Evaluation of the Innovance D-DIMER analytical performance. BiochemiaMedica 2009;19(Suppl 1):S119.
Clinical Institute of Laboratory Diagnosis, Zagreb University School of Medicine and University Hospital Center, Zagreb, Croatia
Corresponding author:desireecoen [at] yahoo [dot] com
Introduction: A widespread use of D-dimer in recent years has led to the development of a number of new fully automated quantitative D-dimer assays.
Methods: We evaluated the analytical performance of the recently developed particle-enhanced immunoturbidimetric assay Innovance D-DIMER on the Behring Coagulation System analyzer (Siemens Medical Solutions Diagnostics).
Results: The within-run coefficients of variation (CVs) for samples with low, borderline, slightly elevated and extremely elevated D-dimer ranged from 2.1% to 5.5%, whereas the between-run CVs for control samples with low and extremely elevated D-dimer ranged from 5.5% to 8.4%. The assay method exhibited very good linearity in the working range between 0.17 and 5.45 mg/L FEU with the analytical detection limit of 0.099 mg/L FEU. The upper reference value determined in 40 plasma samples from apparently healthy volunteers was 0.495 mg/L FEU. For the method comparison study, the results obtained in 419 fresh plasma samples were compared with the results obtained with the Vidas D-DIMER Exclusion on the mini Vidas (bioMérieux). Linear regression analysis according to Passing and Bablok demonstrated a highly significant correlation (y = 1.370x–0.108, r = 0.952, P < 0.001). Bland and Altman difference plots demonstrated slightly higher results obtained with Innovance D-DIMER that were more pronounced with increasing values. Very good agreement between Innovance D-DIMER and Vidas D-Dimer Exclusion was observed in classifying patient results as above or below the cut-off value (kappa coefficient = 0.860; 95% CI, 0.811-0.908).
Conclusions: This study demonstrates that Innovance D-DIMER fulfills all analytical requirements for implementation in daily routine.
Šimek S, Županić D, Pirija M, Percan M, Mujagić R, Honović L.PO3-7: Assessment of diagnostic significance of hematological parametars in discovering the deficiency of vitamin B12 or folate. BiochemiaMedica 2009;19(Suppl 1):S120.
Laboratory of Medical Biochemistry, Pula General Hospital, Pula, Croatia
Corresponding author:danijela [dot] zupanic [at] pu [dot] t-com [dot] hr
Introduction: Megaloblastic anemia is makrocytic anemia (MCV > 97 fL) characterized by nuclear maturation defect and. The deficiency of vitamin B12 or folate is often the reason for that. The aim of this study is to show the relationship between the hematological parametars and the deficiency of vitamin B12 or folate in the group of patients with MCV > 97 fL.
Materials and methods: The study included 69 patients (28 women and 41 men) with MCV > 97 fL and medium age 73 years. Following parametars were measured: hemoglobin concentration, percentage of makrocytes, segment index, percentage of hypersegmented neutrophils (%HS) and the concentration of vitamin B12 and folate. Hematological parameters were determined on cell counter ADVIA 120 (Bayer, USA), %HS and segment index were determined on peripheral blood smear in bright field microscopy (BH-2, Olympus, Japan), and the concentration of vitamin B12 and folate were determined on the analyzer ARCHITECT i2000 SR (Abbott, USA), with CMIA immunochemical method.
Results: The basic criteria for ROC analysis was a diagnostic application of hematological parameters in discovering a deficiency of vitamin B12 or folate. The analysis showed a connection between the deficiency of vitamin B12 and %HS. Calculated area under the curve (AUC) for %HS was 0.656, the confidence interval (95% CI) was 0.529 to 0.769, optimal cut off value was 7% and the value of diagnostic sensitivity and specificity 77.2% and 56.8%.
None of the tested parameters showed diagnostic significance in discovering the deficiency of folate.
Conclusion: With the results of this study we can conclude that the percentage of hypersegmented neutrophils is a sensitive indicator of B12 deficiency in relation to other hematologic parameters.
Miloš M, Coen Herak D, Matišić D, Zadro R.PO3-8: Monoclonal IgM paraprotein with lupus anticoagulant activity – influence on coagulation testing. BiochemiaMedica 2009;19(Suppl 1):S121.
Clinical Institute of Laboratory Diagnosis, Zagreb University School of Medicine and Clinical Hospital Center, Zagreb, Croatia
Corresponding author:marijamilos1 [at] yahoo [dot] com
Introduction: The presence of monoclonal immunoglobulin can induce disturbances of either primary or secondary hemostasis. Hereby we report 3 cases with monoclonal IgM paraprotein and prolonged screening coagulation tests despite absence of bleeding manifestations.
Methods: Routine coagulation tests were performed for all patients: PT with recombinant thromboplastin Innovin and human placental thromboplastin Thromborel S; APTT with Actin FS; one-stage factor assays with the same PT- and APTT-reagents (Siemens Medical Solutions Diagnostics). The presence of lupus anticoagulant (LA) was tested according to the guidelines proposed by the SSC Subcommittee on Lupus Anticoagulant and Phospholipid-Dependent Antibodies of the ISTH. In addition, antibodies against prothrombin and beta-2-glycoprotein I were determined with enzyme immunoassay (AESCU. Diagnostics GmBh). Immunofixation of patients’ sera was performed on Sebia Hydrasys.
Results: Prolonged PT with Innovin and APTT were obtained, without corrections after mixing studies. In factor assays, inhibition of almost all factor activities was detected, with rise in factor activities after multiple dilutions of plasma samples. Sequentially, the presence of LA was confirmed. Suprisingly, PT testing with Thromborel S resulted in normal PT-results, suggesting the possible presence of antibodies against prothrombin. This was confirmed by positive finding of antibodies against both prothrombin and beta-2-glycoprotein I. Based on accidental detection of monoclonal IgM in patient 1 (43.0 g/L), monoclonal IgM was also confirmed in other 2 patients (28.8 g/L and 6.97 g/L, respectively).
Conclusions: This combination of findings could be assigned to the presence of monoclonal IgM paraprotein with both antiprothrombin and anti-beta2-glycoprotein I specificity and strong LA activity.
KutnjakV1, Rogić D2, Kajinić D3, GojčetaK4, Golubić-Čepulić B4. PO3-9: Umbilical cord blood – the importance of hematology analyzer data in estimating the appropriateness of the unit for long term storage. BiochemiaMedica 2009;19(Suppl 1):S122.
1Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia
2Clinical Institute of Laboratory Diagnosis, Zagreb University School of Medicine and Clinical Hospital Center, Zagreb, Croatia
3General hospital Slavonska Po@ega, Slavonska Požega, Croatia
4Zagreb University Hospital, Zagreb, Croatia
Corresponding author:dunjarogic [at] hotmail [dot] com
Uvod: Allogenic or autologous hematopoetic stem cells transplantation is a standard therapy procedure for hematologic malignancies and some other disorders. Recently, umbilical cord blood (UCB) has been increasingly used as an alternative stem cells source. The aim of this study was to assess the accuracy of hematologic analyzers for UCB analysis, and to compare the available methods for hematopoietic progenitor cells number determination.
Materials and methods: 223 samples of UCB were analyzed by flow cytometry and two hematologic analyzers (Sysmex XE-2100 and Abbott Cell Dyne Sapphire). In this study we investigated the possibility of stem cells detection and numbering by hematology analyzer instead of flow cytometry. We also compared hematological analyzers in their ability to detect eritroblasts, which are always present in cord blood and must be detected accurately to avoid falsely elevated number of leukocytes. This is crucial for the accurate feasibility assessment of each unit stored for possible use in stem cell transplantation.
Results and conclusions: The results of our study show that for hematopoetic stem cells detection, the correlation of two methods (hematologic analyzer and flow cytometry) is very low (r = 0.31) and not significant, probably due to the low number of stem cells in UCB. Flow cytometry remains the only recommended method for hematopoetic stem cells detection in cord blood.
Leukocyte count accuracy in UCB – one of the hematologic analyzers (Abbot Cell Dyn Sapphire) performed considerably worse than the other (Sysmex XE 2100) with respect to the ability to detect erytroblasts, when compared to the reference manual method (respective correlation coefficients 0.77 vs. 0.98).