In human and veterinary medicine calprotectin is most widely used in diagnosing different gastro-intestinal diseases. The aim of this study was to assess the stability of canine calprotectin (cCP) in ...serum after storage at low temperatures and imprecision of the method.
Blood samples were collected from dogs with different clinical diagnoses. Twenty-two dogs were included in this study. Calprotectin concentration was measured 4 hours after serum separation (T0), and after being frozen at - 80 °C for 8 (T1) and 16 weeks (T2). The maximum permissible difference (MPD) was derived from the equation for calculating total error (TE) TE = %Bias + (1.96 x %CV), where bias and coefficient of variation (CV) were defined by the manufacturer. The dogs enrolled in this study were patients admitted during the morning (9-12 a.m.), on the day the first measurement was performed. All sample analysis for determination of stability were done in duplicates. For determination of within-run precision, the two patients' serum samples were analyzed in 20 replicates. Imprecision was assessed by analyzing 20 replicates on one plate on two samples where high and low concentrations were anticipated.
The calculated value of MPD was 32.52%. Median calprotectin concentrations were higher at T1 114.08 μg/L (IQR = 55.05-254.56) and T2 133.6 μg/L (IQR = 100.57-332.98) than at T0 83.60 μg/L (IQR = 50.38-176.07). Relative and absolute bias at T1 (49.3%; 45.98 μg/L) and T2 (109.93%; 94.09 μg /L) have shown that calprotectin concentrations increase after long term storage at - 80 °C.
The results of the present study indicate that c-CP was not stable for 16 weeks at low storage temperature (- 80 °C). Considering the observed change in the concentration of c-CP at T1, a storage time of 8 weeks should be safely applied. The method imprecision was not satisfactory, especially in the lower concentration range.
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In 2019 The Croatian Working Group for Laboratory Hematology, on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine, wanted to explore the background in field of ...laboratory hematology routine practice among Croatian laboratories in order to develop future strategies for producing national recommendations, if needed.
During April and May 2019, a comprehensive survey covering all main parts of the total testing process within the field of laboratory hematology among Croatian medical laboratories was conducted. The survey comprised 49 inquiries. Data was collected using Survey Monkey (Palo Alto, CA, USA). All collected data was anonymized.
The response rate was 72%. There is still a substantial number of laboratories that have only three-part differential hematology analyzers (9%). Furthermore, a very high number of laboratories did not perform analyzer verification prior to implementation into routine work (31%). Out of those who have verified their analyzers, a diversity of guidelines and recommendations were used. Nearly 10% of the laboratories do not have a defined policy regarding specimen rejection. The majority of the participants perform internal quality control daily (83%), however, only 51% of respondents evaluate the agreement between different hematology analyzers on daily basis. Although more than 90% of Croatian laboratories have a defined policy regarding specimen rejection, only 61% of respondents continuously monitor quality indicators in routine practice.
The survey revealed substantial differences in all aspects of laboratory hematology practices among Croatian medical laboratories, indicating the need for universal recommendations at the national level.
This study investigated the releasing dynamics of serum ST2 and calprotectin in patients with acute IS. The study included acute IS patients (N = 20) with an NIH Stroke Scale score ≥8. Sampling was ...performed at seven time points: after admission (T0) and at the following 24 h consecutive intervals (T1–T6). Primary outcome at 90 days was evaluated using the modified Rankin scale: 0–2 for good and 3–6 for poor functional outcome. The secondary outcome was all-cause mortality after 90 days. Fifteen patients had a poor outcome, and eight died. Results showed a statistically significant difference in ST2 concentrations between good and poor outcomes at T0 (p = 0.04), T1 (p = 0.006), T2 (p = 0.01), T3 (p = 0.021), T4 (p = 0.007), T5 (p = 0.032), and for calprotectin T6 (p = 0.034). Prognostic accuracy was highest for ST2 at T1 for a cut-off > 18.9 µg/L (sensitivity 80% and specificity 100.0%) and for calprotectin at T5 for a cut-off > 4.5 mg/L (sensitivity 64.3% and specificity 100.0%). Serum ST2 and calprotectin-releasing dynamics showed a valuable prognostic accuracy for IS outcomes.
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The aims of this study were to verify the bile acids (BA) method and to establish reference intervals (RIs) for bile acids (BA) and biochemical and haematological parameters in Croatian pregnant ...women.
BA spectrophotometric method verification was performed on Siemens Atellica Solution CH 930 automated analyser using Sentinel reagent. Stability, precision, trueness, linearity, and RIs, as well as lipemia interference were tested according to CLSI guidelines. BA, biochemical, and haematological parameters were measured in serum (BA, biochemical) and whole blood (haematological) samples of fasting healthy third-trimester pregnant women from Croatia (n=121). The establishment of the RIs was done
according to the CLSI EP28-A3C:2010 guideline. Selected reference individuals' data were analysed using parametric, non-parametric, and robust methods.
Stability study showed that BA are stable in serum samples for 2 days at 20 °C, 14 days at 4-8 °C, and 22 days at -20 °C. The precision study and adult RIs verification met the criteria. Linearity was verified for the concentration range of 3.5-172.1 μmol/L whereas the lipemia interference test showed a positive bias (%) in BA concentration. The determined reference limits generally exhibited better precision for haematological parameters, being lower than the upper recommended value 0.2, unlike biochemical parameters. Haematological parameters showed notable differences between pregnant and non-pregnant women, while many biochemical parameters' RIs remained similar. Only ALT and GGT showed lower non-comparable RI upper limits in the population pregnant women.
Spectrophotometric BA method showed satisfactory performance and all examined parameters were within the set criteria. Moreover, RIs for key biochemical and haematological parameters, including BAs, have been established for the first time in the population of Croatian pregnant women.
Automated erythrocyte sedimentation rate (ESR) analysers are based on different methodology than Westergren method. It is questionable whether ESR values obtained from those analysers are comparable ...with determined values with Westergren method. The aim was verification of the precision, method comparison and accuracy of automated ESR analysers: Roller 20PN (Alifax S.p.A., Polverara, Italy) and iSED (Alcor Scientific, Smithfield, USA).
Blood samples (N = 752 for Roller 20PN and N = 213 for iSED) were sampled into K
EDTA (Kima, Italy) tubes for automated and 3.8% Na-citrate tubes (Kima, Italy) for Westergren method. The data was divided into three groups according to the ESR values obtained with the Westergren method: Group Low (L) (ESR ≤ 20 mm), Group Medium (M) (ESR 21-60 mm), and Group High (H) (ESR ≥ 61 mm). Method agreement was assessed by Bland-Altman analysis and Passing-Bablok regression.
Analyser iSED has shown better comparability with Westergren method (bias 0.0 (95%Cl -1.4 to 1.5) range than Roller 20 PN (bias = - 6.4 (95%Cl - 7.1 to -5.7) in the whole measuring. For Roller 20 PN, Passing-Bablok regression has shown constant and proportional difference for Groups L and M, and for iSED only for Group H. Roller 20 PN had lower sensitivity (0.51 (95%Cl: 0.45-0.57) than iSED (0.72 (95%Cl: 0.59-0.80) while they had comparable specificity (> 0.90) and accuracy (≥ 0.80) in comparison with the Westergren method.
Both analysers are not comparable with the Westergren method and should not be used interchangeably.
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The objective was to determine the associations between several biochemical indicators and the dynamics of concentration change across four physical fitness phases over the period of a competitive ...season. Furthermore, associations between serum calprotectin and biomarkers of inflammation or muscle injury and physical indicators were examined.
Twenty professional male water polo players (median age: 28 (22-42)) were included in this study. Serum creatine kinase activity was determined by the automated photometric UV method. The concentrations of calprotectin, C-reactive protein, and myoglobin were measured using an automated immunoturbidimetric method, while an automated immunochemistry method was employed for interleukin-6, troponin I, and cortisol determination. Tests of repeated strength, maximal strength, and static strength were used to evaluate physical activity.
Serum calprotectin concentrations expressed in median and IQR were significantly different: T1: 2.92 g/mL (2.47; 3.86); T2: 2.35 g/mL (1.26; 2.87); T3: 2.27 g/mL (1.60; 3.27); and T4: 1.47 g/mL (1.04; 2.85) (
= 0.004). Cortisol concentration and CK activity showed significant changes among phases (
= 0.049 and
= 0.014, respectively). Each physical activity examined showed a significant seasonal decrease (all
values were 0.001). Calprotectin serum concentration and indicators of muscular injury, inflammation, and physical activity were found to be correlated during particular stages of the seasonal examination.
Calprotectin values determined throughout one competitive season decreased as training intensity among water polo players increased. Serum calprotectin concentrations and indicators were related to biochemical markers of inflammation and muscle damage.
Testosterone levels in female athletes are increased due to their physical activity and correlate with their exercise volume. We therefore hypothesized that the reference intervals (RIs) derived from ...the general population are not applicable for female athletes. The aim of this study was to evaluate the applicability of the given RIs for 6 commercially available testosterone immunoassays in a group of female athletes.
Our study included 121 female athletes from various sporting disciplines (water polo, handball, volleyball, football, and basketball). The physical activity score was assessed by the Short Form of the International Physical Activity Questionnaire. Total testosterone was measured in serum samples by the reference LC–MS/MS method and six different immunoassays (Abbott Architect 2nd Generation Testosterone, Beckman Coulter Access Testosterone, Roche Elecsys Testosterone II, Siemens Atellica® IM Testosterone II (TSTII), Siemens IMMULITE 2000 Total Testosterone, and Snibe MAGLUMI™ Testosterone).
There were statistically significant differences in age (P = 0.042), weight (P = 0.001), height (P < 0.001), and BMI (P < 0.001) between athletes across different sports. Their quantitative measurements of physical activity and testosterone concentration did not differ significantly between subgroups of various sports, P = 0.167 and P = 0.181, respectively. All immunoassays had a positive absolute and relative bias, in comparison with the LC–MS/MS. The manufacturer’s RI was not verified for Abbott Architect, Beckman Coulter Access, and Roche Elecsys Testosterone methods, with the highest percentage of athletes above RI for Beckman Coulter (30%).
We demonstrated that the upper reference limit provided was too low for some young female athletes. Clinical laboratories should consider implementation of the new proposed RIs.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The aim of this study was to perform a comprehensive verification of a 6-part differential haematology analyser Siemens Advia 2120i (Erlangen, Germany), prior to its routine implementation.
Our ...verification protocol included: precision (within- and between-run), estimated bias (%) as measure of trueness, which was calculated from observed and manufacturers’ declared value, analytical measuring interval (AMI), carryover, confirmation of a limit of blank (LoB), determination of a limit of detection (LoD) and limit of quantitation (LoQ). The K2 ethylenediaminetetraacetic acid (EDTA) patients’ leftover samples were used for verification of analyser Advia 2021i. Acceptance criteria were based on manufacturer technical specifications (Siemens), 2016 state-of-the-art criteria (Vis and Huisman), and EFLM Biological Variation Database.
The within- and between-run precision were acceptable for all parameters and the lowest coefficients of variation were observed for mean corpuscular volume (MCV) (0.3% and 0.6%, respectively). Estimated bias was within the acceptance criteria for all parameters except for MCV (the estimated bias was 2.2% (acceptance criteria 2.0%). AMI was confirmed for all tested parameters (r > 0.99). The carryover estimates ranged from 0.1% for platelet count (Plt) to 0.6% for red blood cell count and were within the manufacturers’ specifications (≤ 1%). Manufacturers’ claims for LoB were confirmed for leukocytes, erythrocytes, haemoglobin, and platelets. The estimated LoD and LoQ were 0.05 x109/L and 0.1 x109/L for white blood cell count, while for Plt values were 2 x109/L and 3 x109/L, respectively.
Analytical performance of the Siemens Advia 2120i meets predefined quality goals and is suitable for routine use in a clinical laboratory.
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