Cytometry Part B: Clinical Cytometry最新文献

{"title":"Central nervous system involvement in multiple myeloma as a first manifestation of relapse and potential diagnostic pitfall.","authors":"Jessy Ranaivomanana, Léa Ryffel, Muriel Newinger, Amira Mejri, Mario Ojeda-Uribe, Pierre De Marini, Bernard Drénou, Magali Le Garff-Tavernier, Thomas Lefebvre, Agathe Debliquis","doi":"10.1002/cyto.b.70035","DOIUrl":"https://doi.org/10.1002/cyto.b.70035","url":null,"abstract":"<p><p>Central nervous system involvement in multiple myeloma (CNS-MM) is a rare complication usually observed in advanced relapse or refractory settings. The detection of malignant plasma cells in the cerebrospinal fluid (CSF) on the basis of morphology and flow cytometry (FCM) confirms this extra-medullary lesion. The records of 459 MM patients treated between 2016 and 2025 were reviewed, identifying four cases with CSF involvement. CSF and bone marrow samples were processed and analyzed on the basis of morphology and FCM using a dedicated 9-color panel to detect malignant plasma cells. Interleukin-6 (IL-6) and interleukin-10 (IL-10) concentrations were also measured in the CSF using Cytometric Bead Assay. Three patients were in complete remission when CSF analysis revealed CNS-MM. Two patients presented no cerebral lesions, although spinal cord compression was observed for one of them. Two patients presented interference in FCM analysis, supporting the penetration of daratumumab into the CSF. Interleukin measures in the CSF showed low or undetectable IL-10 levels but elevated IL-6 concentrations in some cases. Overall prognosis was poor, with the death of three patients and only one remaining alive at 2 years. CNS-MM could be the first sign of relapse among patients previously in complete remission. It should be envisaged in the case of any myeloma patient with unexplained neurological symptoms, even when imaging is negative. Flow-cytometry panels need to be optimized for plasma-cell detection, especially among patients treated with anti-CD38 antibodies. Identifying patients at risk remains crucial, but prognostic biomarkers such as IL-6 still require validation.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147834712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acute myeloid leukemia with mast cell differentiation evolving into mast cell leukemia.","authors":"Liangli Wang, Jie Xu, Lianqun Qiu, Beenu Thakral, Wei Wang, L Jeffrey Medeiros, Hussein A Abbas, Sa A Wang","doi":"10.1002/cyto.b.70036","DOIUrl":"https://doi.org/10.1002/cyto.b.70036","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147834648","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Standardizing stem cell enumeration: A methodological comparison of single and dual flow cytometry platforms.","authors":"Hind Shakah, Alaa Alwahidi, Lara Sarhan, Habibah AlAittan, Saleh Khudirat, Ohood Hammad, Maysa Al-Hussaini","doi":"10.1002/cyto.b.70034","DOIUrl":"https://doi.org/10.1002/cyto.b.70034","url":null,"abstract":"<p><p>Two flow cytometry methods are used for stem cell (CD34⁺) enumeration; single platform (SP) and dual platform (DP). While several studies reported comparable results, others suggested superiority of the SP method. This study evaluated variations between both methods using a modified workflow. A total of 54 fresh and thawed specimens, including mobilized peripheral blood, apheresis products, and umbilical cord blood, were analyzed using both methods. High concordance between SP and DP methods was observed for absolute viable CD34⁺ counts in fresh and thawed specimens (p = 0.088 and 0.427, respectively), as well as for CD34⁺ viability (p = 0.085 and 0.801). Absolute viable WBC counts were comparable between methods in thawed specimens (p = 0.124), whereas a modest statistical variation was observed in fresh specimen group (p = 0.039), largely influenced by umbilical cord blood samples. Variation in absolute viable CD34⁺ counts remained within clinically acceptable limits, with median variations of 2.4 for fresh and 1.4 for thawed samples. SP and DP methods demonstrated high concordance for absolute viable CD34⁺ enumeration and CD34⁺ viability in fresh and thawed specimens. Although a modest variation in viable WBC counts was observed in fresh samples, this did not affect CD34⁺ enumeration and remained clinically acceptable. While SP provides a standardized approach, the DP method offered greater gating flexibility, with fewer technical resources required, and was approximately 70% more cost-effective, supporting its use as a practical alternative in appropriate laboratory settings.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147764669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization and validation of a flow cytometry-based HLA-B27 assay on BD FACSLyric.","authors":"Azzeddine Tahiat, Fethi Meçabih, Houssam Boumzou, Halla Benkortbi, Nabila Attal, Kamel Djenouhat","doi":"10.1002/cyto.b.70031","DOIUrl":"https://doi.org/10.1002/cyto.b.70031","url":null,"abstract":"<p><p>The BD™ HLA-B27 GS145.2-based assay for flow cytometric HLA-B27 typing has been validated on earlier-generation cytometers but not on the increasingly adopted BD FACSLyric platform, highlighting the need for platform-specific validation. While prior transfer strategies relied on conversion of the source instrument's parameters, we performed a de novo analytical validation of the HLA-B27 Kit on the FACSLyric. Our strategy encompassed determination of the assay's gray zone and rigorous evaluation of analytical performance following CLSI H62 guidelines. A total of 125 patient samples were analyzed in parallel by real-time PCR (RT-PCR) and flow cytometry. Sixteen patients (12.8%) were HLA-B27 positive by RT-PCR. A ROC-derived mean fluorescence intensity (MFI) cut-off of 6627 achieved 100% sensitivity and 99.1% specificity. The empirically defined gray zone (6627-7073) was narrow, encompassing only two samples. To strengthen analytical robustness, bootstrapping was applied to derive a 95% confidence interval around the cut-off, yielding an expanded gray zone of 5779-7327. Using this interval, 7 samples (5.6%) were classified as equivocal, comprising four HLA-B27-positive cases and three samples carrying cross-reactive alleles (HLA-B*42, HLA-B*37, and HLA-B*44). Outside the gray zone, flow cytometry showed complete concordance with RT-PCR. MFI values above 7327 ensured unequivocal HLA-B27 positivity with 100% specificity. Precision was high (intra-assay 5.43%, between-run 1.84%, between-day 5.13%), with no carryover and reliable typing up to 24 h post-collection. These results establish a robust, reproducible, and scalable framework for accurate first-line HLA-B27 detection on the FACSLyric platform.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147632839","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"To fret over FRET: Unintended fluorescence energy transfer artifacts and their implications for flow cytometry panel design and interpretation as highlighted by TRBC1/2 assays.","authors":"Hao-Wei Wang, Dana Delgado Colon, Kyle Murphy, Richard R Sochacki, Robert Honec, Jacob Wellek, Isabella Bristol, Jung Jang, Truc Ho, Mariela Monreal, Constance M Yuan","doi":"10.1002/cyto.b.70032","DOIUrl":"10.1002/cyto.b.70032","url":null,"abstract":"<p><p>Förster resonance energy transfer (FRET) serves as the fundamental mechanism underlying a wide array of technologies employed in biomedical research and clinical diagnostic applications. However, unintended FRET is often overlooked, despite its potential to introduce assay artifacts that may lead to misinterpretation. Here, we examine the impact of unwanted FRET on clinical flow cytometric testing, focusing on the T-cell assay incorporating TRBC1 and TRBC2, which has emerged as a powerful method for assessing T-cell clonality. We illustrated the effect using a representative case of spurious antigen expression associated with false TRBC restriction and demonstrate through control experiments that this artifact arises when target antigens in close proximity are labeled with fluorochromes having overlapping spectra. We quantified the spectral overlap integral J(λ), and generated a matrix for commonly used fluorochromes, providing a practical tool to identify fluorochrome pairs with a high FRET propensity that should be avoided when labeling closely situated antigens. Overall, recognizing the potential effects of unintended FRET and implementing strategies to prevent or minimize its impact are essential for ensuring accurate interpretation of clinical flow cytometric assays.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13127340/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147638178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Basophilic blast crisis of chronic myelogenous leukemia presenting de novo: Diagnostic contribution of flow cytometry.","authors":"Chen Glait Santar, Tanya Badelbayov, Dan Benisty, Ben-Zion Katz","doi":"10.1002/cyto.b.70033","DOIUrl":"https://doi.org/10.1002/cyto.b.70033","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147632887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"COMIP-001: A clinical 13-antibody, 12-color flow cytometry panel for optimized immunophenotyping of T/NK cells in human leukemia and lymphoma diagnostics","authors":"Nicole So,&nbsp;Sara Monaghan,&nbsp;Katelynn Davis,&nbsp;Grant Bullock,&nbsp;Wendy Shallenberger,&nbsp;Ahmad Al-Attar","doi":"10.1002/cytob.70015","DOIUrl":"10.1002/cytob.70015","url":null,"abstract":"<p>The objective of this 12-color/13-antibody single-tube panel is to assist in the diagnosis of T/NK-cell leukemias and lymphomas. In the clinical setting, the absence of a standardized T/NK-cell panel limits inter-laboratory data comparability, contributes to diagnostic variability, and results in redundant efforts across laboratories to design and validate panels for flow cytometry. Developing and promoting a panel that allows for rapid and fairly comprehensive labeling of surface T/NK-cell markers, including the anti-T-cell receptor β-chain constant region 1 (TRBC1) antibody, will streamline the workflow by establishing a standard immunophenotyping T/NK-cell panel for detecting neoplastic T/NK cells and lay the foundation for future COMIPs.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"110 2","pages":"73-76"},"PeriodicalIF":2.7,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cytob.70015","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146141351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prognostic impact of immunophenotypic classification for newly diagnosed NPM1-mutated acute myeloid leukemia","authors":"Beatriz Martín-Herreros,&nbsp;Lourdes Cordón,&nbsp;Claudia Sargas,&nbsp;Samuel Romero,&nbsp;María Dolores Linares,&nbsp;Pilar Lloret,&nbsp;Irene Navarro-Vicente,&nbsp;Isabel Cano,&nbsp;Evelyn Acuña-Cruz,&nbsp;Laura Torres-Miñana,&nbsp;Rebeca Rodríguez-Veiga,&nbsp;David Martínez-Cuadron,&nbsp;Blanca Boluda,&nbsp;Eva Barragán,&nbsp;Amparo Sempere,&nbsp;Pau Montesinos","doi":"10.1002/cyto.b.70016","DOIUrl":"10.1002/cyto.b.70016","url":null,"abstract":"<p><i>NPM1</i> mutations are among the most frequent genetic alterations in acute myeloid leukemia (AML) and have been associated with several immunophenotypic features. This study aims to characterize the immunophenotypic profile of this entity at diagnosis using multiparametric flow cytometry and to evaluate the impact of phenotype classifications on prognosis. Immunophenotypic analysis enabled the identification and characterization of various populations previously described in this entity, including immature (CD117⁺ HLA-DR⁺), neutrophil-committed (CD117^{+/heterogeneous} HLA-DR⁻), and monocytic (with variable CD117 expression, HLA-DR⁺, and CD64⁺) leukemic cells, which were identified as leukemic expansions, as well as populations presenting immunophenotypic alterations. Cases were distributed in seven immunophenotypic patterns based on R-classification that considers the presence of the three previously described leukemic cell populations but independently of their proportion and relative distribution. Most of the patients included in this study (30, 33%) exhibited an immature and monocytic leukemic cells population (pattern 4), followed by those patients (16, 18%) with a predominant expansion of immature leukemic cells (pattern 1), and those (16, 18%) characterized by a neutrophil-committed leukemic population (pattern 2). Less frequent patterns included (11, 12%) neutrophil-committed and monocytic cells population (pattern 6), (7, 8%) coexistence of the three populations (pattern 7), (6, 7%) immature and neutrophil-committed leukemic cells population (pattern 5), and (5, 5%) predominant expansion of monocytic leukemic cells population (pattern 3). Our analysis indicated that cases with monocytic leukemic cells population predominance exhibit a trend towards a lower cumulative incidence of relapse compared to other leukemic populations, either as the predominant population or across different immunophenotypic patterns. However, differences did not reach statistical significance. These findings help us to contribute with additional information about the biological and immunophenotypic signature of <i>NPM1-</i>AML.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"110 2","pages":"118-127"},"PeriodicalIF":2.7,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146194229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prospective validation of podoplanin expression as a diagnostic biomarker of acute promyelocytic leukemia","authors":"Camilla Maria de Alencar Saraiva,&nbsp;Carla Roberta Peachazepi Moraes,&nbsp;Bruno Kosa Lino Duarte,&nbsp;Gislaine Borba Oliveira,&nbsp;Herton Luiz Alves Sales Filho,&nbsp;Paula de Melo Campos,&nbsp;Sara Teresinha Olalla Saad,&nbsp;Erich Vinicius De Paula","doi":"10.1002/cyto.b.22252","DOIUrl":"10.1002/cyto.b.22252","url":null,"abstract":"<p>Acute promyelocytic leukemia (APL) is a medical emergency that needs immediate diagnosis and treatment. Podoplanin, a transmembrane glycoprotein that binds CLEC-2 on platelets, was recently demonstrated to be abnormally expressed in leukemic blasts in APL, as opposed to other forms of AML, in a study using thawed primary cells. This study aimed to explore and validate the diagnostic accuracy of measuring podoplanin expression by flow cytometry in the differential diagnosis of APL and other forms of acute myeloid leukemia (AML) as part of the diagnostic work-up of all cases suspected of AML in an academic hematology center. Podoplanin expression was measured by flow cytometry in bone marrow samples obtained at disease presentation from all consecutive adult patients suspected of AML. Results from 24 APL patients were compared with those from 23 non-APL AML patients matched by age and sex. Markedly higher PDPN expression was observed in APL patients when compared to other AML patients, with an area under the curve of 0.92 (95%CI: 0.82–1.0, <i>p</i> &lt; 0.0001) for the percentage of positive cells. Combining an optimal cutoff of 7.66% for PDPN-positive blasts and 1691 for the mean fluorescence index of PDPN expression, APL was identified with a sensitivity of 87.5% and a specificity of 100%. Moreover, PDPN expression presented a negative correlation with platelet count and fibrinogen levels. PDPN expression measured by flow cytometry can accurately differentiate between APL and other forms of AML in a real-world clinical setting, contributing to the diagnosis of this form of acute leukemia. If confirmed in larger prospective studies, the negative association of PDPN expression with fibrinogen and platelet counts supports the concept that this biomarker can potentially contribute to the clinical characterization of APL.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"110 2","pages":"128-133"},"PeriodicalIF":2.7,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22252","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145033111","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"International Clinical Cytometry Society 2023 workload survey of clinical flow cytometry laboratories","authors":"D. Werner,&nbsp;M. A. Linden,&nbsp;L. E. Turner,&nbsp;F. Kreisel,&nbsp;A. Al-Attar,&nbsp;A. Dunlop,&nbsp;A. Ali,&nbsp;T. Denny,&nbsp;W. Kern,&nbsp;V. Litwin,&nbsp;G. Marti,&nbsp;H. Olteanu,&nbsp;C. Trindade,&nbsp;L. Zhang,&nbsp;B. Langworthy,&nbsp;P. K. Wallace,&nbsp;S. A. Monaghan","doi":"10.1002/cyto.b.22259","DOIUrl":"10.1002/cyto.b.22259","url":null,"abstract":"<p>Clinical flow cytometry laboratories are facing rising test volumes, greater assay complexity, and increasing requirements for quality control and assay validation. In response, the International Clinical Cytometry Society (ICCS) conducted a workload survey in early 2023 to gather updated information on assay volumes, complexity, staffing, and technology. Data analysis focused on identifying correlations between length of time to introduce new assays and other factors as a means to gain insight about laboratories that seem to be either adapting or struggling. Flow cytometry assays were categorized into 3 levels of technical/interpretative complexity: high (e.g., measurable/minimal residual disease (MRD assays)), moderate (e.g., leukemia/lymphoma assays (Assays_L&amp;L), excluding MRD assays), and low (e.g., CD4 count). Annual assays per staff member were calculated according to staff involved in case sign-out (Staff_Signout) or other laboratory operations (Staff_LabOps). Respondents were from 101 laboratories in the United States (69.3%), Canada (4.0%), and other countries (26.7%). Low, moderate, and high technical/interpretative complexity assays were performed in 85.1%, 97.0%, and 47.5% of all laboratories, respectively. Median annual total assays (Assays_Total) per laboratory were 3515 and, based on complexity, were 1518.5 (low), 1808.8 (moderate), and 350 (high). Among all laboratories, the median time (interquartile range) to introduce new Assays_L&amp;L was 6 mos. (4–12 mos.), to introduce MRD assays was 11 mos. (5–12 mos.), and to validate/go-live with new cytometers was 8 mos. (4–12 mos.); these times positively correlated with each other. This study confirmed significantly increased workload since the prior ICCS 2013 workload survey with a concurrent decrease in Staff_LabOps. Faster introduction of new assays correlated with other successes, including quicker validation of and going live with new cytometers. Among all laboratories, those that performed myeloid MRD assays versus those that did not were also found to be faster to introduce new assays. The need for sufficient staffing has been emphasized because laboratories with both higher annual volumes of myeloma MRD assays and higher ratios of Assays_Total per Staff_LabOps were slower to introduce new assays. “Lack of staff and/or time dedicated or protected for assay development” and, more generally, “staff number” were the most commonly identified major barriers for new assay development, with the former specifically linked to slower introduction of new assays among all laboratories.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"110 2","pages":"102-117"},"PeriodicalIF":2.7,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145399564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}