Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy

Kaido, Masako; Takagi, Yuri; Kono, Mari; Nakazawa, Fumie; Yamamoto, Shiori; Wada, Atsushi; Morikawa, Takashi

doi:10.1016/j.plabm.2017.05.001

Cited by 10 publications

(18 citation statements)

References 17 publications

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“…The anticoagulants Heparin and EDTA-2K are commonly used during blood collection for blood cell analysis [ 19 ]. We investigated whether there were differences in the measured values of %Teff, %Treg, and K-index using the WB method depending on whether heparin or EDTA-2K was used.…”

Section: Resultsmentioning

confidence: 99%

Establishing a whole blood CD4+ T cell immunity measurement to predict response to anti-PD-1

et al. 2022

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Background Biomarkers that can accurately predict the efficacy of immune checkpoint inhibitors (ICIs) against programmed death 1 (PD-1) ligand in cancer immunotherapy are urgently needed. We have previously reported a novel formula that predicts the response to treatment with second-line nivolumab with high sensitivity and specificity in patients with non-small cell lung cancer (NSCLC) previously treated with chemotherapy. The formula was based on the percentages of CD62LlowCD4+ T cells (effector T cells; %Teff) and CD4+CD25+FOXP3+ T cells (regulatory T cells; %Treg) in the peripheral blood before treatment estimated using the peripheral blood mononuclear cell (PBMC) method. Here, we investigated the applicability of the formula (K-index) to predict the response to treatment with another ICI to expand its clinical applicability. Furthermore, we developed a simpler assay method based on whole blood (WB) samples to overcome the limitations of the PBMC method, such as technical difficulties, in obtaining the K-index. Methods The K-index was evaluated using the PBMC method in 59 patients with NSCLC who received first-line pembrolizumab treatment. We also assessed the K-index using the WB method and estimated the correlation between the measurements obtained using both methods in 76 patients with lung cancer. Results This formula consistently predicted the response to first-line pembrolizumab therapy in patients with NSCLC. The WB method correlated well with the PBMC method to obtain %Teff, %Treg, and the formula value. The WB method showed high repeatability (coefficient of variation, < 10%). The data obtained using WB samples collected in tubes containing either heparin or EDTA-2K and stored at room temperature (18–24 °C) for one day after blood sampling did not differ. Additionally, the performance of the WB method was consistent in different flow cytometry instruments. Conclusions The K-index successfully predicted the response to first-line therapy with pembrolizumab, as reported earlier for the second-line therapy with nivolumab in patients with NSCLC. The WB method established in this study can replace the cumbersome PBMC method in obtaining the K-index. Overall, this study suggests that the K-index can predict the response to anti-PD-1 therapy in various cancers, including NSCLC.

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Section: Resultsmentioning

confidence: 99%

Establishing a whole blood CD4+ T cell immunity measurement to predict response to anti-PD-1

et al. 2022

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“…The NRBC count used in this study was reported as a percentage (%NRBC). NRBC detection in the WNR channel was performed on the Sysmex XN10 hematology analyzer (Sysmex Corporation) by the fluorescent flow cytometric technology using the acidic buffer and polymethine dye, accompanied with optical light scatter detection, which is claimed to reduce interference from lipids and lyse‐resistant red blood cells 19 . Approximately 9530 events per sample are counted, allowing for the lower limit of detection of 0.1% or 0.02 × 10 9 /L as reported by Da Rin G et al 14,20 The hemoglobin levels and WBC count were also collected for further analysis.…”

Section: Methodsmentioning

confidence: 99%

Improving the efficiency of the autoverification workflow for nucleated red blood cell reporting in the hematology laboratory

Chinudomwong

Khongjaroensakun

Chatachote

et al. 2021

Int J Lab Hematology

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Objective Although the microscopic manual count is considered the standard method for NRBC enumeration, modern hematology analyzers can perform this task automatically with reliable accuracy and efficiency. This study aims to evaluate the diagnostic performance of the Sysmex XN hematology analyzer and to construct the optimal workflow for accurate and efficient NRBC reporting. Methods Specimens containing different levels of NRBC were included. Analytical performance was evaluated via method comparison with flow cytometry (FCM) and manual count (MC). Clinical sensitivity was analyzed by ROC analysis using manual count as the standard method. Results Correlation study of %NRBC with FCM and MC demonstrated an r value of 0.925 (95% CI 0.905 to 0.942) and 0.990 (95% CI 0.987 to 0.992) with a mean difference of −0.8 (95%CI: −6.7 to +5.0) and +0.50 (95% CI: −6.7 to +7.7), respectively. When the automated NRBC count was equal to zero and >0.07 × 109/L, the false‐negative rate and false‐positive rate were 100%, respectively; hence, manual slide review could be omitted. A false‐positive rate of 72.7% was noted in specimens containing NRBC count less than 0.07 × 109/L. Conclusion The Sysmex XN can help improve the efficiency of NRBC enumeration owing to its accuracy, rapidity, and automation. However, further studies are required to improve the accuracy of detection in specimens containing a very low level of NRBC.

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“…Sysmex XN-3000 analyzes NRBC in a common WBC & NRBC (WNR) channel using two reagents: Lysercell WNR haemolysing reagent & Fluorocell WNR Nucleic acid dye. 10 Further analysis involves flow cytometric measurements of side fluorescence and forward scatter. Side fluorescence identifies NRBCs by measuring nucleic acid content, while, forward scatter analyses cell size.…”

Section: Methodsmentioning

confidence: 99%

Performance Evaluation of Nucleated Red Blood Cell (Nrbc) Count Using a Fully Automated Haematology Analyzer Versus Manual Counting

Ahmed

Mahmood

Uddin

et al. 2021

PAFMJ

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Objective: To evaluate the performance of Nucleated RBC (NRBC) Count using a fully automated haematology analyzer versus manual counting. Study Design: Cross-Sectional Study. Place and Duration of Study: Department of Hematology, Armed Forces Institute of Pathology, from Sep 2019-Jun 2020. Methodology: Routine fresh whole blood samples were run on Sysmex XN-3000 automated haematology analyzer and 384 samples with results of ≥0.1% Nucleated red blood cells were included in this study. Manual NRBC counting was carried out twice on Leishman-stained peripheral blood smears from all 384 samples. Comparison between manual and automated nucleated red blood cell counting methods was statistically analyzed through linear regression analysis & coefficient correlation. The degree of agreement between two methods was analyzed through Bland-Altman plot. Finally, concordance between the two methods was also analyzed at 5 different ranges of nucleated red blood cells. Results: Linear regression analysis revealed a (r2) value of 0.97. Regression equation was calculated as XN = 0.76MC ± 1.28, with 95% limits of agreement between ± 40.42% and -24.47%. A mean bias of 7.97% was demonstrated through Bland-Altman plot. Concordance analysis revealed a concordance rate of 93.74% (360/384). Nucleated red blood cell counting between two methods were more concordant when nucleated red blood cell counts were <200%. Conclusion: Nucleated red blood cells counting by XN-3000 automated hematology analyzer is statistically comparable to manual nucleated red blood cell counting. We suggest that automated counting can be adopted in routine hematology laboratory as a replacement of manual NRBC counting.

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Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy

Cited by 10 publications

References 17 publications

Establishing a whole blood CD4+ T cell immunity measurement to predict response to anti-PD-1

Establishing a whole blood CD4+ T cell immunity measurement to predict response to anti-PD-1

Improving the efficiency of the autoverification workflow for nucleated red blood cell reporting in the hematology laboratory

Performance Evaluation of Nucleated Red Blood Cell (Nrbc) Count Using a Fully Automated Haematology Analyzer Versus Manual Counting

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