Tokac AGB, Aday A, Erdem S
… +8 more, Bagatir G, Don B, Ozcan O, Yavuz AS, Besisik SK, Nalcaci M, Cefle K, Ozturk S
Int J Lab Hematol
· 2026 May · PMID 42153575
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INTRODUCTION: Chronic myeloid leukemia (CML) is a disease characterized by Philadelphia (Ph) translocations. These translocations can be classical or variant. The structural features and diagnostic implications of varian...INTRODUCTION: Chronic myeloid leukemia (CML) is a disease characterized by Philadelphia (Ph) translocations. These translocations can be classical or variant. The structural features and diagnostic implications of variant Philadelphia translocations remain incompletely defined, and they display considerable cytogenetic heterogeneity. METHODS: In this retrospective study, variant Ph translocations identified by conventional cytogenetic analysis and fluorescence in situ hybridization (FISH) were systematically classified among 639 patients diagnosed with CML. A total of 35 patients with variant Ph translocations were included in the analysis. Molecular follow-up data, when available, were assessed using RT-qPCR analyses in a subset of patients. RESULTS: Chromosome analysis revealed 2 simple and 33 complex variant Ph translocations. FISH analysis, performed in 20 patients, identified deletions involving BCR, ABL1, or both in a limited number of cases. Additional chromosomal abnormalities and secondary translocations accompanied variant Ph translocations in four patients. The partner chromosomes involved in variant Ph translocations showed marked diversity, involving multiple chromosomal loci. CONCLUSION: Variant Philadelphia chromosome translocations in CML exhibit substantial cytogenetic diversity, reflecting the complexity of their underlying genomic architecture. The rarity and heterogeneity of these rearrangements complicate their classification and interpretation in routine diagnostic practice. Descriptive reporting of variant Ph translocations may contribute to a better understanding of their diagnostic complexity and support more accurate cytogenetic interpretation in CML.
Zhang Q, Ma X, Wu Q
… +6 more, Duan Y, Yang C, Tang X, Xu C, Guan M, Wu Z
Int J Lab Hematol
· 2026 May · PMID 42151105
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INTRODUCTION: Current CALR mutation screening in myeloproliferative neoplasms (MPNs) faces a trade-off between coverage and cost: allele-specific qPCR and fragment analysis are inexpensive but cover only part of the vari...INTRODUCTION: Current CALR mutation screening in myeloproliferative neoplasms (MPNs) faces a trade-off between coverage and cost: allele-specific qPCR and fragment analysis are inexpensive but cover only part of the variant landscape, while sequencing costs $300-500 per sample with 3-5 day turnaround. High-resolution melting (HRM) offers both but suffers from subjective interpretation. We developed HRM-CALR-AI, an automated CALR screening platform. METHODS: HRM-CALR-AI was developed on 2667 samples and combines YOLOv8s-cls deep learning with CatBoost meta-classifier for open-set recognition of untrained mutations. Validation included three independent laboratories, 188 consecutive JAK2-negative MPN patients, and a head-to-head comparison with capillary electrophoresis fragment analysis. RESULTS: HRM-CALR-AI processed 96 samples in 90-min at < $10 per test, with limits of detection of 6.25% (CALR-1) and 12.5% (CALR-2) variant allele frequency. On a 480-sample held-out set, it achieved 93.9% accuracy (95% CI: 91.7-95.8), AUC of 0.996 (95% CI: 0.993-0.998), and 89.1% (57/64) recognition of untrained rare variants. Clinical screening identified 160 wild-type (85.1%), 15 CALR-1 (8.0%), nine CALR-2 (4.8%), and four rare variants (2.1%), including a previously unreported variant (c.1145_1158delinsTCCT) classified as CALR-2-like. Multicenter accuracy was 98.1% (ICC 0.868). Against fragment analysis, HRM-CALR-AI showed equivalent detection of length-altering variants (kappa = 0.933) and additionally covered nonlength-altering variants that are invisible to size-based separation. CONCLUSIONS: HRM-CALR-AI combines open-set recognition with low-cost HRM to detect rare and unreported variants at ~50-fold lower cost and same-day turnaround. Multicenter reproducibility and benchmarking against fragment analysis support implementation as a frontline screening tool where NGS access is limited.
Int J Lab Hematol
· 2026 May · PMID 42138564
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INTRODUCTION: RUNX1 is a commonly mutated transcriptional regulator of hematopoiesis in acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Mutated RUNX1 (mRUNX1) may associate with cross-lineage immunopheno...INTRODUCTION: RUNX1 is a commonly mutated transcriptional regulator of hematopoiesis in acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Mutated RUNX1 (mRUNX1) may associate with cross-lineage immunophenotypic aberrancy, presenting potential complications for blast lineage assignment at diagnosis. METHODS: Clinical and laboratory data were reviewed for patients with RUNX1 mutations (N = 125) at Vancouver General Hospital from 2016 to 2022. Diagnostic flow cytometry data were reanalyzed, and RUNX1 mutation characteristics and associations with pathologic, comutation, and flow cytometric features were assessed. RESULTS: Missense RUNX1 mutations (overall 42.9% missense, 57.1% truncating) were enriched in the runt homology domain (RHD; 81.7%, Adj. p = 0.011). Several recurrent hotspot mutations were seen, including common hotspot variants. The most commonly comutated genes with RUNX1 in both acute leukemia with mRUNX1 and MDS-mRUNX1 were ASXL1 (44.4%; 59.3%), SRSF2 (40.7%; 37.3%), and TET2 (35.8%; 55.9%). The mRUNX1 group demonstrated frequent positivity for B-lymphoid markers, with 14.4% having increased expression in at least three B-lineage markers. Differences in antigen expression were associated with RUNX1 mutation type and location, with increased CD10 expression associated with RUNX1 mutations in the RHD (Adj. p = 0.02764), and increased CD79a expression associated with RUNX1 missense mutations (Adj. p = 0.004452). CONCLUSION: In our cohort, we observed missense variants clustering in the RHD and recurrence of common pathogenic variants. Higher-risk comutations were found in a substantial fraction of mRUNX1 cases which may contribute to its adverse risk associations. Comparable degrees of aberrancy in B-lineage markers were seen in mRUNX1 patients versus the comparator cohorts of MPAL-wtRUNX1 and AML with rearranged RUNX1.
Int J Lab Hematol
· 2026 May · PMID 42138489
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Laboratory hematology generates large, complex datasets that increasingly exceed the capabilities of traditional spreadsheet-based analytical workflows. Despite their familiarity, spreadsheets obscure analytical logic, e...Laboratory hematology generates large, complex datasets that increasingly exceed the capabilities of traditional spreadsheet-based analytical workflows. Despite their familiarity, spreadsheets obscure analytical logic, encourage manual data manipulation, and lack durable records of analytical steps, limiting transparency, auditability, and reproducibility. These limitations present specific challenges in clinical laboratory settings subject to regulatory oversight. Script-based data analysis offers a practical alternative by encoding analytical workflows as explicit, ordered, and reusable operations. This review outlines the structural limitations of spreadsheet-centered analysis, illustrates how these constraints contribute to analytical error, and introduces script-based analysis using R as a reproducible framework suited to laboratory hematology. Practical applications are highlighted across method comparison, reference interval estimation, quality control, flow cytometry, and operational monitoring. The discussion also includes the importance of complementary technologies like integrated development environments, version control, and large language models. Adoption of reproducible, script-based analytical tools such as R represents a foundational step toward improving data integrity, analytical rigor, and readiness for advanced analytical techniques in laboratory hematology.
Courville EL, Grant M, Mason E
… +12 more, Chen X, d'Onofrio G, Hedley B, Frater JL, Ouseph MM, Zini G, McFadden S, Han JY, Shirai C, Obstfeld A, Yaeger LH, Pozdnyakova O
Int J Lab Hematol
· 2026 May · PMID 42115681
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OBJECTIVES: Criteria for visual examination of stained peripheral blood smear (PBS) differ among institutions in the United States and internationally. In an effort to standardize review criteria, the International Conse...OBJECTIVES: Criteria for visual examination of stained peripheral blood smear (PBS) differ among institutions in the United States and internationally. In an effort to standardize review criteria, the International Consensus Group for Hematology Review (ICGHR) proposed in 2005 a consensus list of rules for CBC findings that should trigger a review of automated cell counter results and potentially lead to further testing or blood smear review. The primary aim of this paper is to report on the published literature in the past 20 years regarding PBS review criteria and their ability to identify relevant peripheral blood abnormalities. METHODS: We performed a systematic review of the published literature from 2005 to 2025 to investigate and summarize PBS review criteria and performance in the context of automated hematology analyzers in clinical laboratories. RESULTS: Of 5351 citations, 68 studies met our search criteria. These studies included 22 countries and all major hematology analyzer manufacturers. Marked variability was observed in study populations, analyzer flagging criteria, details of PBS visual review, definitions of a "positive" smear, and approaches to statistical data analysis. Across studies, the blast flag sensitivity ranged from 18% to 100% while the blast flag specificity ranged from 17% to 100%. Wide ranges in sensitivity/specificity were also seen for atypical and/or abnormal lymphocyte flags across studies. For studies analyzing the same patient population, less striking variation was seen across instruments. CONCLUSIONS: This systematic review provides a 20-year overview of the literature, highlighting significant variability in PBS review criteria, dependence on study design and hematology analyzer, and the importance of developing harmonized evidence-based guidelines.
Al-Gburi MAA, Safaralizadeh R, Rajabi A
… +1 more, Hosseinpour Feizi M
Int J Lab Hematol
· 2026 May · PMID 42115582
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BACKGROUND: Acute myeloid leukemia (AML) is a clonal illness that is different in both medical and biological traits. Nonetheless, the genetic basis of this malignancy remains inadequately elucidated. MicroRNAs (miRNAs)...BACKGROUND: Acute myeloid leukemia (AML) is a clonal illness that is different in both medical and biological traits. Nonetheless, the genetic basis of this malignancy remains inadequately elucidated. MicroRNAs (miRNAs) are small noncoding RNAs that modify the expression of target mRNAs at both transcriptional and translational levels. In the past decade, miRNAs have evolved a unique mechanism in gene regulation, exhibiting varying expression throughout myeloid differentiation. This investigation examines the serum levels of miR-383, miR-381, and miR-141 in AML and evaluates their diagnostic potential. METHODS: Serum samples from 110 AML patients and 110 normal individuals were evaluated using qRT-PCR. Levels of expression were measured via the 2 technique and normalized by U6. Diagnostic performance was assessed by the ROC curve analysis. RESULTS: MiR-383 was significantly upregulated (p = 0.0001), while miR-381 and miR-141 were downregulated (p = 0.0002 and p = 0.0004, respectively) in AML patients. In addition, a considerable correlation was demonstrated between the expression levels of miR-381 and subtype (p = 0.02). In addition, the expression levels of miR-141 and miR-381 were positively correlated in AML patients (r = 0.99, p ≤ 0.0001). The ROC analysis revealed strong potential of miR-383 (AUC 0.87, 95% CI: 0.82-0.91), miR-381 (AUC 0.71, 95% CI: 064-0.78), and miR-141 (AUC 0.84, 95% CI: 0.79-0.89) as promising diagnostic biomarkers. CONCLUSION: These miRNAs display distinct serum expression levels in AML, with miR-383, miR-381, and miR-141 demonstrating potential as diagnostic biomarkers, necessitating additional confirmation.
Van Vlierberghe M, Louagie H, Ghys T
… +1 more, Degandt S
Int J Lab Hematol
· 2026 May · PMID 42089214
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OBJECTIVES: Haemoglobinopathies are among the most common inherited disorders worldwide. Early detection of heterozygous carriers remains essential for appropriate diagnosis and counselling. The Sysmex 'RBC Defect Workfl...OBJECTIVES: Haemoglobinopathies are among the most common inherited disorders worldwide. Early detection of heterozygous carriers remains essential for appropriate diagnosis and counselling. The Sysmex 'RBC Defect Workflow Optimisation' (RWO) flag is a recently introduced automated tool designed to identify red cell abnormalities suggestive of heterozygous haemoglobinopathies (HGB HTZ), sickle cell disease (SCD), and iron deficiency anaemia (IDA). This study evaluated the diagnostic performance of the RWO flag in routine practice. METHODS: Over 6 months (March-August 2025), 405 samples flagged by RWO (HGB HTZ, SCD, IDA) were prospectively evaluated, and positive predictive values (PPVs) were calculated. In addition, a retrospective analysis of the RWO HGB HTZ, SCD flag was performed on 122 consecutive haemoglobin analyses to assess diagnostic accuracy metrics. Finally, the HGB HTZ β-thalassaemia flag was compared with an external discriminant index, %MicroRBC/%HypoHe ratio (cut-off of 3.7). RESULTS: Prospectively, the Sysmex RWO flag demonstrated PPVs of 81.9% for HGB HTZ, 68.7% for IDA, and 100% for SCD (n = 1). For IDA, PPV increased when analysis was restricted to different exclusion criteria. Retrospective sensitivity for HGB HTZ and SCD was only 25.0%, further clarified in this study. Diagnostic performance of the HGB HTZ β-thalassaemia flag was broadly comparable to that of the %MicroRBC/%HypoHe ratio. CONCLUSIONS: The Sysmex RWO flag represents a feasible and effective screening tool for heterozygous haemoglobinopathies and sickle cell disease in routine clinical practice. Despite modest retrospective sensitivity, its high predictive value and workflow efficiency support its use as a trigger for targeted haemoglobin analysis in everyday laboratory diagnostics.
Chatzikalil E, Delaporta P, Bistas K
… +1 more, Kattamis A
Int J Lab Hematol
· 2026 May · PMID 42083434
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Iron overload is associated with significant health risks, underscoring the importance of understanding its pathophysiology as well as establishing accurate diagnostic and monitoring methods. Chronic iron overload is ass...Iron overload is associated with significant health risks, underscoring the importance of understanding its pathophysiology as well as establishing accurate diagnostic and monitoring methods. Chronic iron overload is associated with either genetic disorders characterized by excessive iron accumulation (hereditary hemochromatosis), or is secondary to diseases of ineffective erythropoiesis and/or requiring regular blood transfusions (like thalassemia, sickle cell disease, myelodysplastic syndromes). Diagnosis is based on clinical suspicion, corroborated by laboratory findings such as elevated serum ferritin and transferrin saturation, along with imaging techniques (magnetic resonance imaging) which facilitate non-invasive assessment of iron levels in parenchymal organs. Elevated ferritin and transferrin saturation and exclusion of secondary causes should prompt genetic evaluation for hereditary disorders predisposing iron overload. Chronic systemic iron overload causes progressive tissue iron accumulation, leading to severe clinical implications, including myocardial dysfunction, liver cirrhosis, and increased risk of hepatocellular carcinoma. Monitoring includes evaluating iron overload indices (serum ferritin, transferrin saturation, liver and heart iron concentration) along with serum and urine indices of parenchymal organ damage at different timepoints regarding the type of disease, patient's age, severity and response to treatment, and aims in improving disease progression and preventing complications. This article provides a comprehensive overview of the pathophysiologic mechanisms and the diagnostic and monitoring techniques of iron overload, in order to revise current knowledge and to raise clinical awareness for effective management.
Int J Lab Hematol
· 2026 May · PMID 42082793
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The direct thrombin inhibitor argatroban is licensed for use in Heparin-induced thrombocytopenia. Original trial data gave the recommendation of monitoring argatroban by Activated Partial Thromboplastin Time (APTT) stati...The direct thrombin inhibitor argatroban is licensed for use in Heparin-induced thrombocytopenia. Original trial data gave the recommendation of monitoring argatroban by Activated Partial Thromboplastin Time (APTT) stating a therapeutic target range of 1.5-3.0 times baseline APTT and not exceeding 100 s. APTT has limitations due to prolongation arising in factor deficiencies, lupus anticoagulants, liver disease, and high FVIII levels leading to potential overestimation of argatroban. Argatroban has demonstrated a plateau effect on APTT at higher concentrations; additionally, APTT reagents have different sensitivity to argatroban, potentially underestimating or overestimating the argatroban. Anti-IIa methods have been recommended as a suitable alternative to accurately quantify argatroban levels. However, there is a lack of consensus on what the target therapeutic range should be. This review will demonstrate how argatroban monitoring by APTT may not be the most suitable method to successfully dose argatroban based on the current state of knowledge and recent published guidelines and highlight the benefits of the anti-IIa methods.
Zini G, Marra J, Rossi E
… +4 more, Bellesi S, Pelliccioni N, d'Onofrio G, Chiusolo P
Int J Lab Hematol
· 2026 May · PMID 42071166
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OBJECTIVES: Digital morphology (DM) systems assisted by artificial intelligence are increasingly being introduced into hematology laboratories; however, data on their performance in routine clinical practice for bone mar...OBJECTIVES: Digital morphology (DM) systems assisted by artificial intelligence are increasingly being introduced into hematology laboratories; however, data on their performance in routine clinical practice for bone marrow aspirates (BMA) remain limited. We evaluated the automated pre-classification generated by a full-field DM system in a large series of BMA samples from patients with onco-hematological disorders. METHODS: We analyzed 350 BMA samples during routine diagnostic activity; they were evaluated using the Siemens Scopio X100 HT system. Automated results were compared with conventional optical microscopy (OM), which served as the reference method for bone marrow differential counts. Results obtained in pre-classification and post-classification were analyzed separately across major cellular lineages and clinically relevant blast thresholds. RESULTS: Sixteen markedly hypercellular samples could not be quantitatively evaluated. In the 334 evaluable BMA samples, the granulocytic series showed very strong correlation between DM and OM at pre-classification (r = 0.85, 95% CI 0.82-0.88), with further improvement after post-classification (r = 0.93, 95% CI 0.91-0.94). Erythroblast percentage showed strong correlation at pre-classification (r = 0.78, 95% CI 0.74-0.82) and very strong correlation after post-classification (r = 0.93, 95% CI 0.92-0.95). Lymphocyte percentage showed moderate correlation at pre-classification (r = 0.55, 95% CI 0.47-0.62) and strong correlation after post-classification (r = 0.78, 95% CI 0.73-0.82). Blast percentage showed strong correlation overall at pre-classification (r = 0.73, 95% CI 0.67-0.77), with very strong correlation in samples with < 5% blasts (r = 0.91, 95% CI 0.88-0.93), but lower correlation in samples with > 5% blasts (r = 0.59, 95% CI 0.48-0.68), improving after post-classification (r = 0.84, 95% CI 0.78-0.88). Low-frequency cell populations and challenging hypercellular smears remained more problematic and required expert morphologic review. CONCLUSIONS: In routine BMA evaluation, the Scopio DM system provided good overall performance for the major marrow lineages, with clear improvement after expert post-classification. Its main value lies in supporting a supervised diagnostic workflow rather than replacing expert microscopic assessment, particularly in challenging samples and in low-frequency or morphologically heterogeneous cell populations.
Pozdnyakova O, Yaeger LH, Shirai CL
… +12 more, McFadden S, Chen X, Obstfeld A, Mason EF, Ouseph MM, Han JY, Zini G, Hedley B, Grant M, D'Onofrio G, Frater JL, Courville EL
Int J Lab Hematol
· 2026 May · PMID 42070993
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Even in the era of modern automated hematology analyzers, visual peripheral blood smear (PBS) review remains an important component of peripheral blood evaluation in a subset of specimens. Despite published PBS review gu...Even in the era of modern automated hematology analyzers, visual peripheral blood smear (PBS) review remains an important component of peripheral blood evaluation in a subset of specimens. Despite published PBS review guidelines, including those issued 20 years ago by the International Consensus Group for Hematology Review (ICGHR), knowledge gaps remain regarding selection and implementation of PBS visual review criteria in clinical laboratories. The purpose of this article, developed by an international working group (the Peripheral Smear Review Working Group [PSRWG]), is to help bridge knowledge gaps by providing practical guidance and items for consideration. PBS review may be indicated for automated analyzer system flags, suspect flags, numerical/definitive flags, or as a component of delta checks. Examples, explanations, and suggestions for each of these categories are provided as are examples of flag modification. Validation of PBS review criteria is a multistep process that includes defining criteria for a positive finding, selecting specimens for study, choosing the slide review method, and determining the statistical approach. Pragmatic approaches and important items to consider are provided for each of these validation steps. PBS visual review workflow must be tailored to the needs of the clinical laboratory/institution. These needs may differ depending on the accrediting agency, laboratory/institutional infrastructure, and population served. A discussion of these variables is provided. In this article, we expand on the proposed ICGHR rules with a discussion of practical considerations in the implementation of PBS review.