Search Results (8)

Background: Reliable platelet (PLT) measurement is crucial for the accurate diagnosis of thrombocytopenia. Several methods exist for automated PLT counting, including the impedance method (PLT-I), as well as optical and fluorescence methods (PLT-F). The impedance method is cost-effective but susceptible to interference from small red blood cells and schistocytes. In contrast, fluorescent assessment offers higher specificity but is more expensive, as it requires additional dyes and detectors. Hybrid platelet counting (PLT-H) combines impedance with measurements from the leukocyte differentiation channel and is available without additional cost. Aim: The aim of this study was to evaluate the accuracy of hybrid PLT counting in anemic samples. Methods: In this retrospective study, PLT counts from 583 unselected anemic samples were analyzed using two different analyzers: the Sysmex XN3500, equipped with fluorescent PLT-F technology, and the Mindray BC6200, which uses both impedance (PLT-I) and hybrid (PLT-H) technologies. Agreement between PLT-I and PLT-F, as well as between PLT-H and PLT-F, was assessed using Bland–Altman plots. Correlation between the methods was evaluated using the Pearson correlation coefficient. Results: The hybrid method demonstrated better accuracy in PLT counting compared to the impedance method. Correlation between PLT-H and PLT-F was excellent, ranging from 0.991 to 0.999. In thrombocytopenic samples (PLT < 50 G/L), the hybrid method also provided more reliable PLT counts than the impedance method, reducing the number of falsely elevated PLT results by nearly fivefold. Conclusions: Hybrid platelet counting yields more accurate results than the impedance method in anemic samples and shows excellent correlation with the fluorescence method. Full article

Objective: Accurate total hemoglobin concentration (ctHb) measurement is critical for clinical decision-making, particularly in acute care, where immediate therapeutic decisions are required. This study evaluated previously established laboratory-based accuracy criteria for ctHb measurements in routine clinical practice at an interdisciplinary operative intensive care unit (IO-ICU), and with particular attention to significantly reduced hemoglobin concentrations. Method: Remaining blood from blood gas analysis (BGA) cuvettes was collected directly at the ICU bedside. From these initial samples, three clinically relevant measurement scenarios were established: direct bedside measurement (Group 01), elevated ctHb levels (Group 02), and lowered ctHb concentrations below 9 g/dl (Group 03). The samples were analyzed using the GEM 4000, GEM 5000 (Werfen GmbH, Muenchen, Germany), ABL90 Flex plus (Radiometer GmbH, Krefeld, Germany), HemoCue Hb 201+, and XN 9000/9100 (Sysmex Deutschland GmbH, Norderstedt, Germany) automatic hematology analyzers. Since each measurement device inherently possesses systematic deviations, no single analyzer was defined as an absolute reference. Instead, the mean value across all tested measurement systems was utilized as a best-fit reference (REF) value. Results: A total of 120 data pairs from 40 ICU patients were analyzed using regression analyses, Bland and Altman (B&A) methods, and tolerance level analysis (TLA). The results demonstrated strong concordance among the evaluated measurement devices across the examined ctHb spectrum (~1–18 g/dL). Moderate systematic deviations identified by B&A analysis were most pronounced at critically low ctHb levels (<6 g/dL). A key outcome was the determination of 95% prediction intervals (PIs), representing a quantifiable range of uncertainties for future bedside measurements. The PIs for Group 03 “low” were in the range of ±7% (relative difference) or ±0.38 g/dL (absolute difference). Conclusion: This study effectively translates previous laboratory findings into clinical practice, highlighting the practical utility of PIs to guide the accurate interpretation of bedside ctHb measurements under acute care conditions. Full article

Cerebrospinal fluid analysis is an important diagnostic test when assessing a neurological canine patient. For this analysis, the total nucleated cell count and differential cell counts are routinely taken, but both involve time-consuming manual methods. To investigate faster automated methods, in this study, the Sysmex XN-V body fluid mode and the deep-learning-based algorithm generated by the Olympus VS200 slide scanner were compared with the manual methods in 161 canine cerebrospinal fluid samples for the total nucleated cell count and in 65 samples with pleocytosis for the differential counts. Following incorrect gating by the Sysmex body fluid mode, all samples were reanalyzed with manually set gates. The Sysmex body fluid mode then showed a mean bias of 15.19 cells/μL for the total nucleated cell count and mean biases of 4.95% and −4.95% for the two-part differential cell count, while the deep-learning-based algorithm showed mean biases of −7.25%, −0.03% and 7.27% for the lymphocytes, neutrophils and monocytoid cells, respectively. Based on our findings, we propose that the automated Sysmex body fluid mode be used to measure the total nucleated cell count in canine cerebrospinal fluid samples after making adjustments to the predefined settings from the manufacturer. However, the two-part differential count of the Sysmex body fluid mode and the deep-learning-based algorithm require some optimization. Full article

In India, where malaria is endemic, the prompt and accurate detection of infections is crucial for disease management and vector control. Our study aimed to evaluate the “iRBC” flag, a novel parameter developed for routine hematology analyzers, for its sensitivity and specificity in detecting Plasmodium vivax (P. vivax) infections. We used residual blood samples from patients with suspected malaria and compared the iRBC flag results with microscopy, which serves as the gold standard. Additionally, we compared the results with rapid immuno-chromatographic tests (RDTs) commonly used in the field. Our study included 575 samples, of which 187 were positive for P. vivax. The iRBC flag demonstrated a high sensitivity of 88.7% and 86.1% on the XN and XN-L hematology analyzers, respectively, and a clinical specificity of 100% on both analyzers. Furthermore, the scattergram derived from each positive dataset exhibited distinct patterns, which facilitated rapid confirmation by laboratory specialists. Notably, the iRBC flag remained effective even in the presence of interfering conditions. Overall, our results indicate that the iRBC flag is a reliable and rapid screening tool for identifying P. vivax in routine blood testing. Our findings have significant implications for malaria detection and control in endemic regions like India. Full article

Background: Tacrolimus (TAC) and mycophenolic acid (MPA) are commonly used immunosuppressive therapies after renal transplant. Our objective was to quantify TAC and MPA concentrations in peripheral blood mononuclear cells (PBMCs) using liquid chromatography tandem mass spectrometry (LC-MS/MS) and to evaluate and validate the performance of the methodology. A prospective follow-up cohort study was conducted to determine whether intracellular concentrations were associated with adverse outcomes in renal transplants. Methods: PBMCs were prepared using the Ficoll separation technique and purified with erythrocyte lysis. The cells were counted using Sysmex XN-3100 and then packaged and frozen according to a 50 µL volume containing 1.0 × 10⁶ cells. TAC and MPA were extracted using MagnaBeads and quantified using an LC-MS/MS platform. The chromatography was run on a reversed-phase Waters Acquity UPLC BEH C18 column (1.7 µm, 50 mm × 2.1 mm) for gradient elution separation with a total run time of 4.5 min and a flow rate of 0.3 mL/min. Mobile phases A and B were water and methanol, respectively, each containing 2 mM ammonium acetate and 0.1% formic acid. Renal transplant recipients receiving TAC and MPA in combination were selected for clinical validation and divided into two groups: a stable group and an adverse outcome group. The concentrations were dynamically monitored at 5, 7, 14, and 21 days (D5, D7, D14, and D21) and 1, 2, 3, and 6 months (M1, M2, M3, and M6) after operation. Results: Method performance validation was performed according to Food and Drug Administration guidelines, showing high specificity and sensitivity. The TAC and MPA calibration curves were linear (r² = 0.9988 and r² = 0.9990, respectively). Both intra-day and inter-day imprecision and inaccuracy were less than 15%. Matrix effects and recoveries were satisfactory. The TAC and MPA concentrations in 304 “real” PBMC samples from 47 renal transplant recipients were within the calibration curve range (0.12 to 16.40 ng/mL and 0.20 to 4.72 ng/mL, respectively). There was a weak correlation between PBMC-C_0TAC and WB-C_0TAC (p < 0.05), but no correlation was found for MPA. The level of immunosuppressive intra-patient variation (IPV) was higher in PBMC at 77.47% (55.06, 97.76%) than in WB at 34.61% (21.90, 49.85%). During the dynamic change in C_0TAC, PBMC-C_0TAC was in a fluctuating state, and no stable period was found. PBMC-C_0TAC did not show a significant difference between the stable and adverse outcome group, but the level of the adverse outcome group was generally higher than that of the stable group. Conclusions: Compared with conventional therapeutic drug monitoring, the proposed rapid and sensitive method can provide more clinically reliable information on drug concentration at an active site, which has the potential to be applied to the clinical monitoring of intracellular immunosuppressive concentration in organ transplantation. However, the application of PBMC-C_0TAC in adverse outcomes of renal transplant should be studied further. Full article

This study investigated the biological effects on circulating monocytes after challenge with SARS-CoV-2 recombinant spike protein. Whole blood collected from seven ostensibly healthy healthcare workers was incubated for 15 min with 2 and 20 ng/mL final concentration of recombinant spike protein of Ancestral, Alpha, Delta, and Omicron variants. Samples were analyzed with Sysmex XN and DI-60 analyzers. Cellular complexity (i.e., the presence of granules, vacuoles and other cytoplasmic inclusions) increased in all samples challenged with the recombinant spike protein of the Ancestral, Alpha, and Delta variants, but not in those containing Omicron. The cellular content of nucleic acids was constantly decreased in most samples, achieving statistical significance in those containing 20 ng/mL of Alpha and Delta recombinant spike proteins. The heterogeneity of monocyte volumes significantly increased in all samples, achieving statistical significance in those containing 20 ng/mL of recombinant spike protein of the Ancestral, Alpha and Delta variants. The monocyte morphological abnormalities after spike protein challenge included dysmorphia, granulation, intense vacuolization, platelet phagocytosis, development of aberrant nuclei, and cytoplasmic extrusions. The SARS-CoV-2 spike protein triggers important monocyte morphological abnormalities, more evident in cells challenged with recombinant spike protein of the more clinically severe Alpha and Delta variants. Full article

The early identification of septically infected newborn infants is important for ensuring good outcomes. Blood cell differentiations are helpful, but they are often time consuming and inaccurate. In this study, we evaluated the use of automatic white blood cell differentiations by flow cytometry for the diagnosis of neonatal sepsis. Episodes of suspected infection in neonates were retrospectively classified into two groups, unlikely infection (UI, levels of Interleukin-6 < 400 pg/mL or CRP within 48 h < 10 mg/L), n = 101 and probable infection (PI, Interleukin-6 ≥ 400 pg/mL or CRP within 48 h ≥ 10 mg/L), n = 98. Complete blood cell counts were performed by Sysmex XN-9000^® using flow cytometry. Relative and absolute proportions of immature granulocytes were evaluated. Unexpectedly, the absolute count of immature granulocytes was significantly lower in the group of PI compared to UI neonates. Similar results were found when analysing the relative proportion of immature granulocytes among all neutrophil granulocytes. On the other hand, manually counted immature to total (I/T) ratios of granulocytes were higher in PI than in UI infants. Therefore, we conclude that differentiations of granulocytes by Sysmex XN-9000^® can be used to distinguish between infected and uninfected neonates if the results are interpreted according to our findings. A low count of immature granulocytes as determined by Sysmex XN-9000^® may indicate neonatal infection. Full article

The performance of platelet (PLT) counting in thrombocytopenic samples is crucial for transfusion decisions. We compared PLT counting and its reproducibility between Mindray BC-6800Plus (BC-6800P, Mindray, Shenzhen, China) and Sysmex XN-9000 (XN, Sysmex, Kobe, Japan), especially focused on thrombocytopenic samples. We analyzed the correlation and agreement of PLT-I channels in both analyzers and BC-6800P PLT-O mode and XN PLT-F channel in 516 samples regarding PLT counts. Ten thrombocytopenic samples (≤2.0 × 10⁹/L by XN PLT-F) were measured 10 times to investigate the reproducibility with the desirable precision criterion, 7.6%. The correlation of BC-6800P PLT-I and XN PLT-I was arranged moderate to very high; but the correlation of BC-6800P PLT-O and XN PLT-F was arranged high to very high. Both BC-6800P PLT-I vs. XN PLT-I and BC-6800P PLT-O vs. XN PLT-F showed very good agreement (κ = 0.93 and κ = 0.94). In 41 discordant samples between BC-6800P PLT-O and XN PLT-F at transfusion thresholds, BC-6800P PLT-O showed higher PLT counts than XN-PLT-F, except the one case. BC-6800P PLT-O exceeded the precision criterion in one of 10 samples; but XN PLT-F exceeded it in six of 10 samples. BC-6800P would be a reliable option for PLT counting in thrombocytopenic samples with good reproducibility. Full article