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    Eosin-5′-maleimide (EMA)-binding assay as a diagnostic method of hereditary spherocytosis
    (De Gruyter, 2025) Pepeler, Mehmet Sezgin; Falay, Mesude; Aydın, Mürüvvet Seda; Parmaksız, Ayhan; Yılmaz Keskin, Ebru; Alanoğlu, Güçhan; Fettah, Ali; Özet, Gülsüm
    Objectives Erythrocyte membrane disorders are caused by a deficiency of structural proteins in the erythrocyte membrane. Accurate differential diagnosis within this group of disorders (is essential for appropriate management. The eosin-5′-maleimide (EMA) binding assay is a novel test that is used for the differential diagnosis of erythrocyte membrane disorders. In this study, we have examined and reported blood counts, reticulocyte indices, and the EMA binding assay results with clinical findings of cases admitted to our laboratory for suspected red blood cell (RBC) membrane disorder. Methods We performed the EMA binding assay on the blood samples of 103 patients who were screened for hereditary erythrocyte membrane disorders at the Flow Cytometry Laboratory of Ankara Numune Training and Research Hospital. The total cohort was grouped as patients with hereditary spherocytosis (HS) (n=36) and control group (patients without erythrocyte membrane disorders (n=60), and non-HS patients with a preliminary diagnosis of hemolytic anemia (n=7). The control group included during data collection, the results of the EMA binding assay were recorded along with demographic features, clinical information, morphologic features, blood count parameters, RBC and reticulocyte indices, and a conventional osmotic fragility (OF) test. Receiver Operating Characteristics (ROC) analyses were performed to evaluate the diagnostic accuracy of the EMA binding assay and reticulocyte parameters. Results Both EMA testing and flow cytometric (FC) OF test were significantly lower in overall (n=36), ≤10-year-old (n=12), and >10-year-old (n=24) patients with HS than in healthy controls (p<0.001). The EMA binding assay had 100 % sensitivity and specificity in screening HS. Conclusions Combined with conventional blood tests, clinical findings, and medical history, the EMA binding assay is a reliable and convenient tool for screening for HS and differentiating hereditary erythrocyte membrane disorders.
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    A novel differential diagnosis algorithm for chronic lymphocytic leukemia using immunophenotyping with flow cytometry
    (Elsevier, 2023) Narlı Özdemir, Zehra; Falay, Mesude; Parmaksız, Ayhan; Genç, Eylem; Beyler, Özlem; Güneş, Ahmet Kürşad; Ceran, Funda; Dağdaş, Simten; Özet, Gülsüm
    Introduction: The availability of a clinical decision algorithm for diagnosis of chronic lymphocytic leukemia (CLL) may greatly contribute to the diagnosis of CLL, particularly in cases with ambiguous immunophenotypes. Herein we propose a novel differential diagnosis algorithm for the CLL diagnosis using immunophenotyping with flow cytometry. Methods: The hierarchical logistic regression model (Backward LR) was used to build a predictive algorithmfor the diagnosis of CLL, differentiated from other lymphoproliferative disorders (LPDs). Results: A total of 302 patients, of whom 220 (72.8%) had CLL and 82 (27.2%), B-cell lymphoproliferative disorders other than CLL, were included in the study. The Backward LR model comprised the variables CD5, CD43, CD81, ROR1, CD23, CD79b, FMC7, sIg and CD200 in the model development process. The weak expression of CD81 and increased intensity of expression in markers CD5, CD23 and CD200 increased the probability of CLL diagnosis, (p < 0.05). The odd ratio for CD5, C23, CD200 and CD81 was 1.088 (1.050 - 1.126), 1.044 (1.012 - 1.077), 1.039 (1.007 - 1.072) and 0.946 (0.921 - 0.970) [95% C.I.], respectively. Our model provided a novel diagnostic algorithm with 95.27% of sensitivity and 91.46% of specificity. The model prediction for 97.3% (214) of 220 patients diagnosed with CLL, was CLL and for 91.5% (75) of 82 patients diagnosed with an LPD other than CLL, was others. The cases were correctly classified as CLL and others with a 95.7% correctness rate. Conclusions: Our model highlighting 4 markers (CD81, CD5, CD23 and CD200) provided high sensitivity and specificity in the CLL diagnosis and in distinguishing of CLL among other LPDs.

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