Int J Lab Hematol
· 2026 Jun · PMID 42309506
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INTRODUCTION: Although AI-powered digital morphology analyzers are widely used for leukocyte classification, their reliability in detecting clinically significant abnormal cells requires rigorous validation. We evaluated...INTRODUCTION: Although AI-powered digital morphology analyzers are widely used for leukocyte classification, their reliability in detecting clinically significant abnormal cells requires rigorous validation. We evaluated the Mindray MC-80 system by comparing its results with those of conventional hematology analyzers and manual microscopy to assess accuracy, sensitivity, and diagnostic limitations. METHODS: We analyzed 1188 routine blood samples (881 normal, 307 abnormal). Preclassification results from the MC-80 were compared with hematology analyzer results for normal samples and with manual microscopy by senior morphologists for abnormal samples. Inter-method agreement was assessed by Passing-Bablok regression, and sensitivity and specificity were calculated using expert-reviewed findings as the reference standard. RESULTS: The MC-80 preclassification demonstrated excellent correlation with reference assays, with r values greater than 0.9 for the majority of leukocyte subsets. Overall specificity reached over 99% for all cell populations. Excluding blasts and reactive lymphocytes, the sensitivity for all remaining cell types was higher than 90%. For clinically critical abnormal cells, the system attained 75% sensitivity for plasma cells (≥ 1%) and over 90% for the other five abnormal cell groups. Its specificity was above 90% for all abnormal cells except blasts, which had a specificity of 86.73%. Expert review further increased the correlation coefficients by 0.31% to 5.71%. CONCLUSION: The MC-80 provides robust performance for routine leukocyte classification, with a screening capability for most abnormal cells comparable to that of manual microscopy. However, its reduced sensitivity for reactive lymphocytes, blasts, and plasma cells indicates that it cannot fully replace manual review. Targeted optimization is therefore warranted before full integration into diagnostic workflows.
Ferrari C, Coulleray B, Kerneves P
… +8 more, Geara C, Naguib D, Levallet G, Battersby E, Chantepie S, Repessé Y, Nectoux C, Maitre E
Int J Lab Hematol
· 2026 Jun · PMID 42298870
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INTRODUCTION: Quantification and monitoring of NPM1 mutations represent a key tool in the management of acute myeloid leukemia (AML), offering strong prognostic value and serving as a marker of minimal residual disease (...INTRODUCTION: Quantification and monitoring of NPM1 mutations represent a key tool in the management of acute myeloid leukemia (AML), offering strong prognostic value and serving as a marker of minimal residual disease (MRD). Digital PCR (dPCR) provides absolute quantification with high sensitivity and specificity, offering advantages over conventional qPCR-based methods, particularly for rare or atypical NPM1 mutations. METHODS: We adapted and evaluated a digital PCR strategy including both monoplex and multiplex assays targeting the most common NPM1 mutations (types A, B, D) as well as rare variants. A touch-down PCR protocol was optimized to harmonize the dPCR protocol and enhance specificity. Assay performance was evaluated in terms of limit of detection (LoD), limit of quantification (LoQ), linearity, and specificity, and compared to quantitative PCR (qPCR). RESULTS: The multiplex dPCR assay allowed detection of all clinically relevant NPM1 mutations with an LoD of 0.01%, while monoplex type A, B and D assays reached 0.001% for type A, B and D. The method showed strong linearity (R > 0.99 for all targets) and high concordance with qPCR (R = 0.9967). In patient samples, dPCR demonstrated improved sensitivity over qPCR, detecting low-level NPM1 mutations in samples previously negative by qPCR, and enabled early relapse detection. CONCLUSION: The approach enabled early relapse prediction in selected patients and improved molecular follow-up, even for rare NPM1 mutations not covered by standard qPCR panels. It simplifies laboratory routine and improves MRD assessment, according to current ELN guidelines and the growing need for personalized molecular monitoring in AML.
Assadi-Gazvini C, Li C, Arnautou P
… +2 more, Hejl C, Amiot Q
Int J Lab Hematol
· 2026 Jun · PMID 42285774
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We report a 67-year-old man with extranodal NK/T-cell lymphoma, nasal type, revealed by hemophagocytic lymphohistiocytosis. He presented with compressive aerodigestive symptoms and cytopenias. Diagnosis was confirmed by...We report a 67-year-old man with extranodal NK/T-cell lymphoma, nasal type, revealed by hemophagocytic lymphohistiocytosis. He presented with compressive aerodigestive symptoms and cytopenias. Diagnosis was confirmed by bone marrow and lymph node biopsy. Treatment combined etoposide and pembrolizumab-based immunochemotherapy.
Correani A, Niccoletti B, Pavani M
… +5 more, Sampaolo M, Finaurini L, Pelloso M, Montagnana M, Moretti M
Int J Lab Hematol
· 2026 Jun · PMID 42267648
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INTRODUCTION: HemoScreen is an image-based point-of-care (POC) hematology analyzer that performs complete blood counts (CBC) from small blood volumes with minimal handling. These features are particularly relevant in ped...INTRODUCTION: HemoScreen is an image-based point-of-care (POC) hematology analyzer that performs complete blood counts (CBC) from small blood volumes with minimal handling. These features are particularly relevant in pediatrics; however, comparative evidence with central laboratory analyzers in children remains limited. METHODS: We compared HemoScreen POC with Sysmex XN-1000 analyzer in 141 venous K-EDTA samples from pediatric inpatients aged 2 months-17 years. The primary outcome was analytical agreement for white blood cells (WBC), red blood cells (RBC), and platelets (PLT). Secondary outcomes included: (1) analytical agreement for hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), and mean platelet volume (MPV); and (2) diagnostic agreement for WBC, HGB, MCV, and PLT categories. Analytical agreement was evaluated using Passing-Bablok regression, Bland-Altman analysis, and proportions exceeding total allowable error (TEa) thresholds; Spearman's correlation was used to assess the strength of association between methods. Diagnostic agreement was quantified with Cohen's κ before and after Passing-Bablok alignment. RESULTS: Correlations were strong to excellent, with no significant deviation from linearity. Mean biases were below the minimum acceptable limits for most parameters. Using the Passing-Bablok regression as the reference line, the proportion of measurements exceeding limits was below 10% for WBC, RBC, PLT, and most other CBC indices in at least one of the tested TEa thresholds. Diagnostic agreement was excellent for leukopenia, leukocytosis, thrombocytopenia, anemia, and elevated HGB; good-excellent for thrombocytosis and macrocytosis; and good for microcytosis. CONCLUSION: In venous samples from pediatric inpatients (2 months-17 years), HemoScreen showed high analytical and diagnostic agreement with the Sysmex XN-1000, supporting its use in children's hospitals.
Velasco-Rodríguez D, Marcos-Jubilar M, Martínez-Alfonzo I
… +19 more, Martos L, Aguilar C, Vilalta N, Llobet D, Marco-Rico A, Aragoneses D, Marco P, Ricca L, Ferreira D, Lopes S, Mariano-Pego J, Bernardo Á, Moscardó A, Bonanad S, Gassiot S, Gutiérrez-Jomarrón I, Páramo JA, Llamas-Sillero P, Reverter JC
Int J Lab Hematol
· 2026 Jun · PMID 42264461
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INTRODUCTION: Fully automated thrombin generation (TG) assays facilitate the transition from research to clinical application. We conducted a multicenter inter-laboratory study using the ST Genesia analyzer to evaluate a...INTRODUCTION: Fully automated thrombin generation (TG) assays facilitate the transition from research to clinical application. We conducted a multicenter inter-laboratory study using the ST Genesia analyzer to evaluate assay performance and establish reference intervals in healthy adults from Spain and Portugal. METHODS: In this multicenter study, plasma samples from healthy adult volunteers were analyzed across 12 laboratories following a harmonized protocol. TG was measured using the STG-ThromboScreen reagent in the presence and absence of thrombomodulin (TM). Analytical performance and reference intervals were assessed. RESULTS: Within-run and intra-day precision were excellent (coefficient of variation < 10%) for all TG parameters except velocity index. Normalization reduced interlaboratory variability for amplitude-based parameters such as endogenous thrombin potential (ETP) and peak height, whereas its effect was less pronounced for velocity index. Satisfactory interlaboratory performance (|z| ≤ 2) was achieved in 96%-98% of results, and normalized parameters showed minimal bias across centers. In 252 healthy adults, median normalized values were: ETP 131% (IQR 118-142), peak height 116% (IQR 104-127), lag time 2.6 min (IQR 2.3-2.8), and time-to-peak 5.5 min (IQR 5.0-5.9). Median ETP inhibition in the presence of TM was 47.2% (IQR 35.1-61.8). Age had a modest influence on TG parameters, whereas sex differences were observed. CONCLUSIONS: This multicenter interlaboratory study in the Iberian Peninsula demonstrates high reproducibility of TG measurement using the ST Genesia platform with the STG-ThromboScreen reagent. These findings support its use for standardized TG assessment and provide reference intervals for clinical and research settings.
Wang Y, Geng S, Chen X
… +10 more, Huang L, Huang X, Luo Q, Zeng L, Wu P, Zhang Y, Liu K, Du X, Weng J, Lai P
Int J Lab Hematol
· 2026 Jun · PMID 42253149
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INTRODUCTION: This study aimed to characterize T follicular helper (Tfh) cell differentiation and related molecular profiles in chronic-phase chronic myeloid leukemia (CML-CP) patients with distinct molecular responses a...INTRODUCTION: This study aimed to characterize T follicular helper (Tfh) cell differentiation and related molecular profiles in chronic-phase chronic myeloid leukemia (CML-CP) patients with distinct molecular responses and to explore immune correlates of treatment outcomes. METHODS: Peripheral blood CD4 T cells were analyzed in 125 CML-CP patients (8 newly diagnosed, 71 MMR, and 46 pre-MMR) and 22 healthy donors. We assessed CD4 T subset distribution and expression of Tfh-related molecules including Bcl6, Blimp1, PD-1, PD-L1, and PD-L2. RESULTS: Newly diagnosed and pre-MMR patients exhibited decreased CD4 T cell proportions compared to healthy donors and MMR patients (p < 0.05), while pre-MMR patients had elevated frequencies of CD4CXCR5PD-1 Tfh cells and increased PD-1 expression on CD4 T cells (p < 0.05). Bcl6, PD-1, and PD-L1 were upregulated in pre-MMR cases, whereas Blimp1 and PD-L2 remained unaltered. CONCLUSION: Pre-MMR CML-CP is accompanied by aberrant Tfh cell differentiation and an immunosuppressive immune signature linked to suboptimal molecular responses. Targeting immune checkpoints or modulating Tfh cell homeostasis may help improve therapeutic outcomes in CML-CP.
Mishra S, Kumar K, Kumar D
… +3 more, Kamak A, Singh L, Kumar N
Int J Lab Hematol
· 2026 Jun · PMID 42252205
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INTRODUCTION: Tyrosine kinase inhibitors (TKIs) have transformed the treatment of chronic myeloid leukemia (CML); yet, diverse molecular responses and resistance persist. BCR::ABL1 kinase-domain (TKD) mutations constitut...INTRODUCTION: Tyrosine kinase inhibitors (TKIs) have transformed the treatment of chronic myeloid leukemia (CML); yet, diverse molecular responses and resistance persist. BCR::ABL1 kinase-domain (TKD) mutations constitute just a fraction of this resistance, and the impact of additional somatic mutations on disease progression and early molecular response remains incompletely defined. METHODS: This single-centre cohort study analyzed 109 NGS-tested patients with CML, comprising 44 with TKI-resistant disease and 65 newly diagnosed patients. Targeted next-generation sequencing using a 135-gene myeloid panel was performed on 109 patients. An additional pilot subgroup of 30 TKI-resistant patients underwent BCR::ABL1 kinase-domain analysis by PCR/Sanger sequencing and was analyzed separately. Molecular response was assessed using BCR::ABL1 transcript levels on the International Scale and interpreted according to ELN 2020 recommendations. RESULTS: Somatic mutations were identified in 52.3% of TKI-resistant and 29.2% of newly diagnosed patients. All Cohort 1 blast-crisis patients were mutation-positive, and several concurrent abnormalities were more common in Cohort 1 than in Cohort 2, indicating clonal complexity. In Cohort 2, MMR was achieved in 28/39 (71.8%) mutation-negative and 6/13 (46.2%) mutation-positive patients. Mutation-positivity at baseline was associated with reduced MMR chances but not statistically significant (odds ratio 0.34; 95% confidence interval 0.09-1.23; p = 0.099). ASXL1 emerged as the most common non-ABL1 mutation but was not statistically significant. CONCLUSIONS: In this Indian CML cohort, somatic mutations were prevalent in TKI-resistant disease, linked to advanced phase and clonal complexity, and demonstrated a non-significant trend toward lower early MMR at diagnosis, highlighting the importance of genomic testing in this context.
Novelli C, Gatti A, Ierna F
… +2 more, Riccardi E, Cuppari I
Int J Lab Hematol
· 2026 May · PMID 42206535
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INTRODUCTION: Lupus anticoagulant (LA) testing is essential, albeit complex, in the laboratory diagnosis of antiphospholipid syndrome (APS). Given the multi-step workflow and the variability introduced by anticoagulant t...INTRODUCTION: Lupus anticoagulant (LA) testing is essential, albeit complex, in the laboratory diagnosis of antiphospholipid syndrome (APS). Given the multi-step workflow and the variability introduced by anticoagulant therapy, reagent differences, and interpretive approaches, result interpretation requires expert evaluation. To address these challenges, we implemented a middleware-based automated algorithm in our four-hospital institution using the HemoHub system, guided by the latest ISTH recommendations. The objectives were to automate reflex testing, standardize interpretation, and support clinical decision-making. METHODS: The algorithm incorporated rules aligned with good laboratory practice for LA diagnostics, including reflex testing, result interpretation, internal comments, integration of historical data, and auto-validation of negative cases. Retrospective validation was performed on 190 historical cases in which the diagnosis was made conventionally and corroborated by follow-up data. A subsequent prospective blind comparison with manual operator-based workflow was conducted on 481 routine samples. RESULTS: Concordance between automated and manual interpretations reached 100% for LA negative and positive cases, with slightly lower agreement for first-time positives (88.6% and 82.4%). When automated interpretation was not assignable, the system still provided useful internal comments. Additionally, 58.4% of samples were auto-validated, significantly reducing manual workload. Time comparison demonstrated substantial savings for both experienced and less-experienced pathologists. CONCLUSION: The implementation of the automated algorithm improved consistency, reduced interpretation time, and minimized intra- and inter-laboratory variability. By integrating clinical context and historical data, it enhanced diagnostic accuracy. These findings support the use of middleware-based, rule-driven interpretation as a reliable and efficient approach to standardizing LA testing and optimizing laboratory workflow.
Sanfilippo S, Buisson L, Delabie B
… +3 more, Le Bellego S, Dujaric ME, Donnet T
Int J Lab Hematol
· 2026 May · PMID 42185000
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BACKGROUND: Acquired hypofibrinogenemia is frequently associated with major bleeding and requires prompt correction. Rapid and accurate quantification of fibrinogen is essential to guide hemostatic therapy and improve pa...BACKGROUND: Acquired hypofibrinogenemia is frequently associated with major bleeding and requires prompt correction. Rapid and accurate quantification of fibrinogen is essential to guide hemostatic therapy and improve patient outcomes. The qLabs FIB system provides fibrinogen results within minutes at the bedside, supporting timely diagnostic assessment and therapeutic decision-making. This study aimed to validate the device analytical performance in a prospective, observational, multicenter setting by comparing whole-blood qLabs FIB results with lab-based plasma fibrinogen. METHODS: Fibrinogen concentrations from 463 citrated whole-blood samples were measured in parallel using the qLabs FIB and the Clauss method across 8 clinical sites. RESULTS: Overall, a strong correlation was observed between the qLabs FIB and the Clauss laboratory method (r = 0.94). The median of relative differences did not differ significantly between clinical sites. Receiver operating characteristic analysis showed sensitivities and specificities in the 90% range for identifying fibrinogen concentrations below the clinical decision thresholds of 1.5 and 2.0 g/L. CONCLUSION: This study demonstrated a strong agreement of the qLabs FIB to the standard Clauss assay while providing rapid, near-patient fibrinogen measurements. This multicenter study confirms the initial performance of the system from a larger number of samples. The integration of qLabs FIB testing into routine clinical practice could support earlier, more targeted hemostatic interventions, thereby improving the safety and effectiveness of bleeding management.
Albayrak H, Ay ME, Tombak A
… +4 more, Çevik K, Ay Öİ, Kabasakal T, Erdal ME
Int J Lab Hematol
· 2026 May · PMID 42167790
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BACKGROUND: Circadian clock disruption has emerged as a relevant axis in cancer; however, the expression patterns and diagnostic relevance of BMAL1 and CLOCK in multiple myeloma (MM) remain insufficiently defined. METHOD...BACKGROUND: Circadian clock disruption has emerged as a relevant axis in cancer; however, the expression patterns and diagnostic relevance of BMAL1 and CLOCK in multiple myeloma (MM) remain insufficiently defined. METHODS: BMAL1 and CLOCK mRNA expression was quantified by RT-qPCR in bone marrow samples from 46 newly diagnosed MM patients and 13 healthy controls. Group comparisons, ROC analyses, multivariate logistic regression, partial correlation, and machine-learning models (logistic regression, random forest, and support-vector machine) with stratified 5-fold cross-validation (including class-weighted sensitivity analysis) were performed. RESULTS: BMAL1 expression was significantly higher in MM than in controls (1.12 ± 0.33 vs. 0.85 ± 0.33, p = 0.016; Cohen's d = 0.83), whereas CLOCK showed no significant univariate difference (p > 0.2). BMAL1 demonstrated moderate diagnostic performance (AUC = 0.729), while CLOCK alone showed limited discrimination (AUC = 0.617). Combined analysis modestly improved performance (AUC = 0.756). In multivariate logistic regression, BMAL1 remained a strong independent predictor of MM (OR = 3343.4, p = 0.0046), whereas CLOCK showed an inverse adjusted association (OR = 0.0037, p = 0.019). BMAL1 and CLOCK expression were strongly correlated in MM (ρ ≈ 0.80), persisting after adjustment for age, ISS stage, and β2-microglobulin (partial r = 0.847, p = 2.2 × 10-13). Among classifiers, the random forest achieved the best performance (AUC = 0.832; AUPRC = 0.952). Specificity was limited across classifiers, indicating that these performance estimates are exploratory and require validation in larger cohorts. CONCLUSION: MM is characterized by BMAL1 upregulation within a preserved but imbalanced BMAL1/CLOCK axis rather than uniform shifts across circadian genes. Although diagnostic performance is moderate, consistent signals across statistical and machine-learning approaches support an MM-associated circadian expression pattern warranting further validation.
Androulakis N, Nioti E, Dilintas A
… +1 more, Kalpadakis C
Int J Lab Hematol
· 2026 May · PMID 42159456
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Borderline prolongation of routine clotting assays-particularly the activated partial thromboplastin time (aPTT)-is a common interpretative challenge in laboratory hematology. These reproducible but mildly prolonged resu...Borderline prolongation of routine clotting assays-particularly the activated partial thromboplastin time (aPTT)-is a common interpretative challenge in laboratory hematology. These reproducible but mildly prolonged results often lie just beyond the upper reference limit and can trigger unnecessary follow-up, delays, or misinterpretation. Conventional tools such as 1:1 mixing studies may appear corrective due to dilutional masking of weak inhibitors, residual anticoagulants, or low-grade lupus anticoagulant (LA) activity, especially in assays with limited sensitivity. In our laboratory, borderline prolongations prompt a structured, hypothesis-driven workflow that escalates interpretative resolution in sequential steps. We begin with verification of analytical validity and exclusion of confounders (e.g., DOACs, pre-analytical errors), then apply conventional mixing studies interpreted via quantitative correction indices (e.g., Rosner, percent correction). When results remain indeterminate or discordant, we escalate to enhanced mixing ratios (e.g., 4:1, 1:4), followed by factor assays and dilutional analysis including parallelism assessment. Here, we outline an integrated decision-support framework for investigating borderline aPTT prolongation. The framework aligns traditional and enhanced mixing strategies with factor-level interpretation into a coherent, stepwise pathway featuring explicit stop-go points. It is intended for use in advanced or referral coagulation laboratories and emphasizes interpretative rigor over binary thresholds. By optimizing existing tools within a layered interpretative structure, this approach seeks to reduce false reassurance and enhance clarity in borderline aPTT interpretation.