Establishment of Method and Verification for Leukocyte Differential Count of Sysmex Hematology Analyzer / 现代检验医学杂志

Objective To calibrate the leukocyte differential counts and compare between all sysmex hematology analyzers, making the results accurate and reliable.Methods Used the manual visual microscope method as a reference method for leu-kocyte differential count to detect the fresh blood and then did assignment.The target value,calibrating the leukocyte differ-ential counts of Sysmex XE-2100,XS-800i and XN-20 (A1)with the manual value fresh blood,was calibrated and then com-pared hematology analyzers with another fresh blood.Results The leukocyte differential counts of all hematology analyzers were within the 95% confidence interval of the manual visual microscope method.Comparing with the reference system,the correlation coefficient(r)of NEUT%,LYM%,MON%,EOS% and BASO% of XS-800i analyzer were 0.99,0.99,0.98, 0.98 and 0.59,respectively.The correlation coefficient (r)of NEUT%,LYM%,MON%,EOS% and BASO% of XN-20 (A1)analyzer were 0.99,0.99,0.97,0.98 and 0.55,respectively.XS-800i and XN-20 (A1)NEUT%,LYM%,MON% and EOS% were within confidence interval 100%.XS-800i BASO% were within confidence interval 95%.XN-20 (A1)BASO%were within confidence interval 90%.Conclusion This scheme could meet the WS/T 246-2005 demands.In order to ensure the accuracy and the results of mutual recognition of the leukocyte differential counts,the comparison analysis of the results of the manual visual microscope method and hematology analyzer were made.

Predictive value of leukocyte differential count in patients with acute cerebral infarction / 医学研究生学报

Objective Inflammation response is involved in the whole pathological process of acute cerebral infarction ( ACI) , but few reports are seen on its clinical implication in ACI patients .The purpose of this study was to investigate the predictive value of the differential count of leukocytes for stroke severity and early clinical outcomes in the acute phase of cerebral infarction . Methods We collected the clinical and laboratory data of 635 patients diagnosed with ACI within 72 hours of symptom onset and eval-uated the association between the differential count of peripheral blood leukocytes and stroke severity at admission and within 3 days af-ter admission as well as the clinical outcomes at discharge .The neural function impairment scores of the patients were obtained with The NIH Stroke Score ( NIHSS) at admission and on the third day after admission , and the therapeutic results evaluated with the modi-fied Rankin Scale ( mRS) , mRS >2 as poor prognosis .Analyses were performed on the correlation of the differential count of leuko-cytes with NIHSS and mRS scores and its influence on the ACI patients . Results At discharge , the mRS related influencing factors included the total count of leukocytes (OR=1.147, 95% CI:1.038-1.268), count of neutrophil cells (OR=1.227, 95% CI:1.00-1.369 ), count of lymphocytes ( OR =0.508, 95% CI:0.342-0.753), and neutrophil to lymphocyte ratio (NLR) (OR=1.150, 95%CI:1.008-1.314).the NIHSSs were correlated with the counts of leucocytes (r=0.078, P=0.024), neutrophil cells (r=0.083, P=0.019), and lymphocytes (r=0.010, P=0.004) at admission, and with the counts of leucocytes ( r =0.238, P <0.001), neutrophil cells (r=0.335, P<0.001), lymphocytes (r=-0.269, P<0.001), and NLR (r=0.423, P<0.001) on the third day after admission. Conclusion In the acute phase of cer-ebral infarction , the differential count of leukocytes and NLR are valuable for predicting the severity of neurologic impairment and early poor functional outcome .

"Leukocyte differential count in peripheral blood ""with"" single tube/ten colors""flow cytometry" / 中华检验医学杂志

Objective To explore the values of potential clinical application ofsingle tube/ten colorsflow cytometry for leukocyte differential count in peripheral blood.Methods Utilizing multiple monoclonal antibody combinations and the vavious logical gating strategies,the single tube/12 antibodies with no-wash method for the leukocyte differential count in peripheral blood were determined by using 10 colors flow cytometry.Leukocyte differentials of 142 peripheral blood samples were determined by both Beckman-Coulter LH750 hematology analyzer and 10 colors flow cytometry.The results were then compared to standard microscopic examination as a reference method.The clinical diagnostic efficiency ofsingle tube/10 colorsflow cytometry was calculated.The correlations between standard microscopic cytology,single tube/10 colorsflow cytometry and the hematology analyzer were determined.In addition,the clinical diagnosis efficiency for blast counts ofsingle tube/10 colorswere compared to the results determined by BD FACS Calibur flow cytometer.Results The leukocyte differentials were correlated well between the single tube/10 colorsflow cytometry and standard microscopic cytology(r>0.700,P<0.01) except for basophils.The correlations with neutrophilic granulocytes,lymphocytes,immature granulocytes and blasts were superior(r=0.972,0.951,0.801,0.912,respectively,P<0.01).When 1% was selected as the cut-off point for immature granulocytes determined by standard microscopic cytology,the sensitivity and the specificity ofsingle tube/10 colorsflow cytometry were 92%(57/62) and 79% (63/80),respectively.When 0.5% was selected as the cut-off point for blasts detected by standard microscopic cytology,the sensitivity and the specificity were 99% (67/68) and 92% (68/74).Using the immunophenotyping results from BD FACS Calibur as a standard,the sensitivity for detecting blasts bysingle tube/10 colOrsflow cytometry was 100% (40/40),the specificity was 91% (10/11),the positive predictive value was 98% (40/41),the negative predictive value was 100% (10/10) and the accuracy was 98% (50/51).Conclusions Thesingle tube/10 colorsflow cytometry has a excellent correlation with the standard microscopic cytology when applied on leukocyte differential count in peripheral blood.It may potentially use as a subsequent method for verification of abnormal results of complete blood cell count in the future.

Differential Blast Counts Obtained by Automated Blood Cell Analyzers / 대한진단검사의학회지

BACKGROUND: Automated blood cell analyzers often read leukemic blasts as normal cells. In this study, we evaluated the 5-part differential patterns of blasts using automated analyzers to determine if they can differentiate among blast types. METHODS: Blood samples containing 10% or more blasts were collected from patients with acute leukemia (N=175). The 5-part differential count was conducted using DxH 800 (Beckman Coulter, USA) and XE-2100 analyzers (Sysmex Co., Japan), and the results were compared with manual differential counts, which was used as a reference method. RESULTS: The DxH 800 reported the 5-part white blood cell differential count in 98.9% of the cases. The XE-2100 provided an invalid automated differential count in 72% of the cases. Both analyzers counted most lymphoblasts as lymphocytes and most myeloblasts as monocytes. In 11 cases, the DxH 800 reported a 5-part differential count without a blast flag. CONCLUSIONS: Some automated analyzers are able to recognize and count blasts according to their characteristic cell types. Therefore, complete blood counts obtained automatically can provide valuable data for making provisional decisions regarding the lineage of leukemia cells before further investigation.

CELL-DYN Sapphire Hematology Analyzer Performance Evaluation on Leukocyte Differential Counts / 임상검사와정도관리

BACKGROUND: Recent advances of hematology analyzers have improved performance of leukocyte differential counts and have reduced work load of clinical hematology laboratories. We evaluated CELL-DYN Sapphire (Abbott Diagnostics, Santa Clara, CA, USA) performance on leukocyte differential counts according to Clinical and Laboratory Standards Institute (CLSI) document H20-A. METHODS: We evaluated imprecision (short term imprecision from duplication of 147 patients' sample and long term imprecision from three level commercial controls) and accuracy (n=462) of leukocyte differential counts of CELL-DYN Sapphire and compared with those of Sysmex XE-2100 (TOA Medical Electronics Co., Kobe, Japan), ADVIA 120 (Bayer Diagnostics, Tarrytown, NY, USA) and Beckman Coulter LH 750 (Beckman Coulter, Miami, FL, USA). RESULTS: The imprecision of CELL-DYN Sapphire for neutrophils and lymphocytes differentials was low with coefficients of variation (CV) from 1.4 to 6.2%, but the imprecision for basophils was high with CV from 34.7 to 79.6%. The correlation with manual count was good in samples without flags (n=314), with the exception of basophils (r: neutrophils, 0.921; lymphocytes, 0.921; monocytes, 0.653; eosinophils, 0.869; basophils 0.272). The correlation with other hematology analyzers was high except basophils (r: neutrophils, 0.969-0.986; lymphocytes, 0.986-0.990; monocytes, 0.787-0.887; eosinophils, 0.881-0.962; basophils 0.086-0.327). CONCLUSION: The performance on leukocyte differential counts of CELL-DYN Sapphire is comparable to Sysmex XE-2100, ADVIA 120 and Beckman Coulter LH 750. In regards of enumeration of basophils, the comparison with manual counts and other hematology analyzers shows poor agreement.

Analysis of the Bone Marrow Aspirates with Automated Hematology Analyzer for Assessment of the Bone Marrow Cellularity and Effective Hematopoiesis / 대한진단검사의학회지

BACKGROUND: Microscopic examination of the bone marrow (BM) smear has been a major method for the diagnostic and post-therapeutic evaluation of hematologic disease but is laborious and imprecise due to small number of cells counted. Recently, automated reticulocyte counting is available by the automated hematology analyzer. We analyzed the bone marrow aspirates using Coulter GEN S (GEN S) automated hematology analyzer and compared the results with those by the microscopic examination. METHODS: Total nucleated cells (TNC), leukocyte subpopulations, red cell count, hemoglobin and reticulocyte indices of the peripheral blood (PB) and the BM aspirates, were measured by GEN S in 392 samples including 142 normal control samples. Differential counts by microscopic examination of Wright stained BM films were used as a reference. RESULTS: TNC of the BM obtained by automated hematology analyzer correlated with the BM cel-lularity estimated by microscopic examination (r=0.587, P=0.000). The differential counts of neutrophils and monocytes correlated between these two methods (r=0.582, P=0.000, r=0.309, P=0.000). In acute leukemia, TNC of the PB and the BM, and the BM lymphocyte fraction were increased and the BM neutrophil fraction was decreased. In chronic myelogenous leukemia, TNC of the PB and the BM were high but distribution of leukocyte subpopulations was normal. In normal control group, the number of erythroid precusors correlated with the percentages of reticulocyte in the PB (r=0.425, P=0.000), and in patients with increased erythropoiesis, it showed strong correlation with immature reticulocyte fraction (IRF) of the PB (r=0.708, P=0.033). In aplastic anemia, IRF of the PB was reversely correlated to hemoglobin level, but in myelodysplastic syndrome, reticulocyte indices of the PB and the BM had no correlation with hemoglobin level. In patients with increased erythropoiesis, the percentages of reticulocyte in the PB were increased and those of the BM were decreased in proportion to reduction of hemoglobin level in the PB. CONCLUSIONS: Analysis of the BM aspirates using automated hematology analyzer will be useful in screening of pathological hematologic diseases and in estimating the bone marrow cellularity objectively before those by the microscopic examination. In anemia, this study could provide an additional information to evaluate the ineffective hematopoiesis using reticulocyte indices of the PB and the BM.

Significance of Serum Cortisol and Peripheral Blood Leukocyte Differential for the Early Differential Diagnosis of Acute Chest Pain Syndrome

OBJECTIVES: The stress response involves the activation of the hypothalamic-pituitary-adrenal(HPA) axis and the sympathetic nervous system. Corticosteroids have been clearly demonstrated to cause anti-inflammatory and/or immnosuppressive effects in man including granulocytosis in part by decreasing migration into tissue, especially damaged tissues(myocardium), and circulating relative lymphocytopenia. To test whether automated measurements of the the increased serum cortisol-induced hematologic changes in the leukocyte differential significance or not in the initial differential diagnosis of acute myocardial infarction in acute chest pain syndromes. METHODS: 101 consecutive patients with myocardial infarction or myocardial ischemia presenting to the emergency room of Seoul Adventist Hospital with acute chest pain from January 1993 to August 1995(Retrospective group) and from December 1995 to March patients compatible with exclusion criteria in myocardial infarction were excluded. We measured automated leukocyte differential and serial CK-MB level in both groups, and the intial serum cortisol levels in prospective infarction group. RESULTS: 1) Total leukocyte and granulocyte counts were increased in acute myocardial infarction(p<0.01). 2) In acute myocardial infarction group, lymphocyte counts were slightly increased(p<0.05), but relative lymphocytes percentage more significantly decreased(p<0.01). 3) Serum cortisol levels are significantly raised early in the course of the acute myocardial infarction and prior to the elevation of the specific cardiac enzymes on the basis of analytic results of prospective infarction group. 4) Cortisol-induced changes in leukocyte differential were noted with time passes into reverse approximately 4 days later in our study. 5) The leukocyte differential does not shows significant changes in the retrospective myocardial ischemia group, so we arrive in careful conclusion that serum cortisol level seems does not increase. 6) No sexual differences were noted in leukocyte differential. CONCLUSIONS: The serum cortisol level and cortisol-induced leukocyte differential are helpful for initial differential diagnosis of acute myocardial infarction in acute chest pain sysdrome.