Neutrophil scattering data driven pre-microscopic flagging of acute leukemic cases

ABSTRACT Background: The hematology analyzer, Sysmex XN-1000, generates white blood cell count with varying scattering intensities during a complete blood count (CBC) analysis. Objectives: The objectives of the study were to study the predictive role of median and coefficient of variation of neutrophil scattering items in blood samples for differentiation of leukemic subjects. Methods: We evaluated six neutrophil scattering parameters: neutrophil side scatter mean intensity, neutrophil side fluorescence light (SFL) mean intensity, neutrophil forward scatter mean intensity, neutrophil side scatter area distribution width (NE-WX), neutrophil SFL area distribution width (NE-WY), and neutrophil forward scatter area distribution width (NE-WZ), measured in white blood cell differential scattergram generated by the hematology analyzer (Sysmex XN-1000) at an academic medical center. Results: We collected 433 blood samples from acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL) cases and normal controls. AML group showed highly significant differences in the mean values compared with the control group. Out of six neutrophil scattering items, NE-WX, NE-WY, and NE-WZ showed high efficiency, with area under the curve (AUC) values of 0.764, 0.748, and 0.757, respectively, to differentiate AML from ALL cases and control groups. When comparing combined acute leukemia cases (AML plus ALL) with the control group, NE-WX, NE-WY, and NE-WZ generated highly significant AUC values (0.840, 0.884, and 0.801, respectively). Conclusion: The neutrophil scattering parameters generated during CBC analysis provide a new tool for the prediction of acute leukemia and its lineage.

Cell Population Data NE-SFL and MO-WX From Sysmex XN-3000 Can Provide Additional Information for Exclusion of Acute Promyelocytic Leukemia From Other Acute Myeloid Leukemias: A Preliminary Study

No abstract available.

Automated detection of malaria with haematology analyzer sysmex xe-2100.

Background: Diagnosis of malaria is usually made by microscopy [Giemsa, Acridine Orange (AO), and Quantitative Buffy Coat (QBC) assay], which requires expertise. Currently, automated haematology analyzers are being used for complete blood count (CBC), in all acute febrile and non-febrile illnesses which simultaneously detects malaria. The normal scattergram by the analyzer (Sysmex 2100) comprises of five parameters i.e. lymphocytes (pink), monocytes (green), neutrophils (blue), eosinophils (red) with a space between the neutrophil and eosinophil populations. Aims : We carried out a prospective study to compare the efficacy of Sysmex XE-2100 (Sysmex Corporation, Kobe) for detection of malaria in comparison to other conventional techniques. Materials and Methods : 430 cases were analyzed for malaria by microscopy (QBC, AO, Giemsa), ICT (Immunochromatography) and flowcytometric analyzer (Sysmex XE-2100). The abnormal scattergrams were observed as double neutrophil, double eosinophil, grey zone, extended neutrophil zone with a decrease space between eosinophil and neutrophil, and a combination of above patterns. Results : Out of 70 positive cases [49/70 (70%) P. vivax, 18/70 (25.7%) P. falciparum, and 3/70 (4.2%) both P. vivax and P. falciparum], 52 showed abnormal scattergrams by the analyzer. The sensitivity and specificity of hematology analyzer found to be 74.2% and 88%, respectively. Conclusion : Flowcytometric analyzer is a rapid, high throughput device which needs less expertization for the diagnosis of malaria. Hence, it can be used in the diagnostic laboratories as an early modality for diagnosis of malaria in suspected as well as clinically in apparent cases.

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.

Current Status and Proposal of a Guideline for Manual Slide Review of Automated Complete Blood Cell Count and White Blood Cell Dfferential / 대한진단검사의학회지

BACKGROUND: Manual slide review (MSR) is usually triggered by the results of automated hematolgy analyzers, but each laboaratory has different ciriteria for MSR. This study was carried out to investigate the current status of MSR criteria of automated complete blood cell count (CBC) and white blood cell (WBC) differential results and to propose a basic guideline for MSR. METHODS: Total 111 laboratories were surveyed regarding MSR using questionnaires. The questionnaire asked: kinds of automated hematology analyzers used and the presence of criteria triggering MSR in seven categories: 1) CBC results, 2) 5 differential WBC counts, 3) 3 differential WBC counts, 4) automated reticulocyte counts, 5) delta check, 6) instrument flags (or messages), 7) clinical information (wards or diseases). Based on the survey results, we determined basic and extended criteria for MSR. With these criteria, we consulted nine hematology experts to get a consensus. RESULTS: All 111 laboratories had their own MSR criteria. Among 111 laboratories, 98 (88.3%) used more than three criteria for MSR including CBC results and 5-part WBC differential count results and 95 (85.6%) had criteria of flags triggering MSR. For MSR criteria with numeric values, the 10th, 50th, and 90th percentiles of upper and lower threshold values were obtained. The basic guideline for MSR was made. CONCLUSIONS: We proposed a basic guideline for MSR. This guideline would be helpful to hematology laboratories for their daily operation and providing more rapid and accurate CBC and WBC differential results.

Del hemograma manual al hemograma de cuarta generación / From blood manual blood fourth generation

El hemograma o cuadro hemático es una de las pruebas que más se solicita al laboratorio clínico, y sin duda alguna, la prueba de laboratorio que más aporta al clínico en la evaluación de un paciente. Desde el punto de vista técnico se reconocen seis tipos de hemograma, que van desde los tradicionales que se hacen con métodos manuales hasta los más sofisticados que se hacen con métodos electrónicos que utilizan una combinación detecnologías. Se establecen los criterios que definen los tipos de hemograma y se analizan los parámetros desde el punto de vista metodológico, los valores de referencia, las indicaciones clínicas y los aspectos críticos de cada parámetro de acuerdo con la metodología utilizada.El médico debe solicitar el hemograma que le permita tener mayor certeza analítica posible en los parámetros reportados y el laboratorio clínico debe hacer la inversión tecnológica que le permita ofrecer resultados lo más precisos y exactos posible. Palabras clave: hemograma, cuadro hemático, recuento de células, laboratorio, utilidad clínica, valores de referencia.

Evaluation of the Abbott Cell-Dyn Sapphire Hematology Analyzer / 대한진단검사의학회지

BACKGROUND: The performance of Cell-Dyn Sapphire (Abbott Diagnostic, USA) was compared to the Bayer Advia 2120 (Bayer Diagnostics, USA), Sysmex XE-2100 (Sysmex Corporation, Japan), and reference microscopy. METHODS: Three hundred samples for routine CBC and WBC differentials were randomly chosen for a comparison analysis. The Cell-Dyn Sapphire system was evaluated according to the linearity, imprecision, inter-instrument correlations, and white blood cell differential. RESULTS: The CBC parameters (WBC, RBC, hemoglobin and platelet) showed a significant linearity with correlation coefficients greater than 0.99 (P<0.0001). Coefficients of variation (CV) for withinrun and differential count of WBC were less than 5% except for Total CV for monocytes, eosinophils, and basophils and within-run CV for low valued eosinophils. The correlation coefficients with manual count were lower in monocytes, eosinophils, and basophils than in neutrophils and lymphocytes. The correlation with other hematology anlayzers was significant exclusive of basophils. CONCLUSIONS: These results demonstrate that the Cell-Dyn Sapphire has a good linearity, an acceptable reproducibility, a minimal carryover, and a comparable performance with the sysmex XE-2100 and Advia 2120.

Avaliação dos critérios de liberação direta dos resultados de hemogramas através de contadores eletrônicos / Evaluation of the direct liberation criteria of hemogram results from electronic counters

Os aparelhos de automação em hematologia oferecem alta sensibilidade e precisão na realização do hemograma na rotina do laboratório de hematologia. Para avaliar os critérios de liberação dos hemogramas por interfaceamento direto entre os contadores eletrônicos e o setor de emissão dos laudos, foram examinadas 247 lâminas de hemogramas. Entre essas, 149 lâminas foram consideradas concordantes com os critérios de liberação por interfaceamento direto enquanto 98 foram consideradas não concordantes pelos contadores automatizados Sysmex SE 9500 e Cell Dyn 4000. Dentre os hemogramas concordantes, a microscopia sem acesso aos dados numéricos mostrou alteração em apenas uma lâmina, a qual era clinicamente irrelevante. Por outro lado, entre os 98 hemogramas não concordantes, 32 apresentavam alterações. Os dados encontrados comprovam a satisfatória sensibilidade dos sistemas de automação analisados. As lâminas dos hemogramas não concordantes foram reexaminadas, com o objetivo de avaliar os resultados eletrônicos. Esse fato permitiu a análise das causas das reprovações com sugestões para eventuais simplificações do sistema. O presente trabalho mostrou que os resultados emitidos pelos contadores automáticos analisados apresentam uma alta correlação com a avaliação microscópica. Conclui-se, portanto, que os resultados dos hemogramas liberados pelos aparelhos analisados são satisfatórios e estes aparelhos são confiáveis para a rotina no laboratório de hematologia.

Automated equipment used in the hematological laboratory forroutine hemograms offers high sensitivity and great accuracy. Inorder to evaluate the liberation criteria of hemograms by directinterfacing between the electronic counters and the emission ofresults, 247 hemogram slides were examined. Of these using theSysmex SE 9500 and Cell Dyn 4000 automated counters, 149slides were considered concordant with liberation criteria bydirect interfacing while 98 were considered non-concordant. Ofthe concordant hemograms, microscopy without access to thenumeric results showed alterations in only one slide, which wasconsidered clinically irrelevant. On the other hand, of the 98non-concordant hemograms, 32 showed alterations. The resultsdemonstrate the satisfactory sensitivity of the automatic systemsanalyzed. The slides with the non-concordant hemograms werere-examined, in order to evaluate the electronic results. Thus, itwas possible to analyze the causes of the non-concordance andgive suggestions for possible simplifications of the system. Thisstudy shows that the results emitted by the assessed automaticcounters presented a high level of correlation with the microscopycount. Therefore, it can be concluded that the results of thehemograms liberated by the devices are satisfactory and that thesedevices are reliable for the routine use within the hematologicallaboratory. Rev. bras. hematol. hemoter. 2004; 26(3):159-166.

Evaluación del desempeño de contadores hematológicos / Performance assessment of blood cells analyzers

El objetivo de esta presentación es brindar, en forma didáctica y actualizada, una orientación para la evaluación de contadores hematológicos. Primeramente se hace referencia a los niveles de evaluación, la información preliminar referida al instrumento sobre requerimientos de espacio y servicios y al manual de instrucción. En cuanto a las etapas de evaluación se tratan: a) la planificación técnica que incluye acuerdos previos con el fabricante y la planificación de recursos internos; b) calibración, calibradores y materiales de control; c) reactivos; d) muestras: su manipulación y registros de datos; e) ensayo preliminar: normas de seguridad (mecánicas, eléctricas, microbiológicas, químicas, ambientales) e instalación y mantenimiento; f) evaluación del desempeño: efecto de la dilución, coincidencia, precisión intra e interensayo, deriva, evaluación del arrastre, comparabilidad con datos obtenidos por procedimientos de rutina, exactitud, efecto del envejecimiento de la muestra, sensibilidad para muestras anormales, interferencias, análisis de datos y utilidad clínica; g) evaluación de la eficiencia. Finalmente, se sacan conclusiones sobre la base de un ejemplo de evaluación de un equipo de apertura-impedancia

Clinical Usefulness of the Hematopoietic Progenitor Cell Counts in Predicting the Optimal Timing of Peripheral Blood Stem Cell Harvest

Although enumeration of CD34+ cells in the peripheral blood (PB) on the day of apheresis predicts the quantity of those cells collected, the flow cytometric techniques used are complex and expensive, and several hours are required to obtain the result in the clinical practice setting. The Sysmex SE-9000 automated haematology analyzer provides an estimate of immature cells, called hematopoietic progenitor cells (HPC). The aim of this study was to evaluate the clinical usefulness of HPC in predicting the optimal timing of peripheral blood progenitor cells (PBPC) harvest. Studies were performed on 628 aphereses from 160 patients with hematologic or solid malignancies. Spearman's rank statistics was used to assess correlation between HPC, WBC, mononuclear cells (MNC), and CD34+ cells. A receiver operating characteristic (ROC) curve was drawn for cutoff value of HPC, and predictive values of the chosen cutoff value of HPC for different target CD34+ cell collections were calculated. The PB HPC had a stronger correlation (rho=0.592, por=1 x10(6)/kg with sensitivity of 75%. Positive and negative predictive values of HPC >or=50x10(6)/L for CD34+ cells >or=1x10(6)/kg were 59.7% and 81.1%, respectively. In the clinical practice setting, applying variable cutoff values of HPC would be a useful tool to predict the optimal timing of PBPC collection.

Evaluation of the stability of Cell Dyn 16 Tri Level as a control material for laboratory external quality assessment scheme on hematology.

The implementation of a laboratory test should always implement a laboratory quality control program, i.e internal quality control and external quality assessment. In an external quality assessment scheme, a control material that is stable over delivery until tested by the participating laboratory. In this study, we evaluated the stability of Cell Dyn 16 Tri Level (TL) control material at room temperature (26-32 degrees C), stored in a transport vessel containing ice pack, and the precision and accuracy of the instrument Cell Dyn 1400. The control used was Cell Dyn 16 TL with low value (L), normal value (N) and high value (H). This study was done in the Clinical Pathology Department of FKUI-RSCM during February 2001 until May 2001. Control material was stored room in a transport vessel containing ice pack for 15 days, then analysed macroscopically, microscopically and evaluated for its stability. Test for precision and accuracy was done within run and for precision between day on Cell Dyn 1400. The result of this study showed a macroscopic change beginning on day 14 (L) day 12 (N) and day 15 (H). Microscopic change was observed on day 13 (L and N) and day 15 (H), Erythrocyte and hemoglobin level was stable until day 15. Changes in leukocyte was seen on day 14 (L), day 12 (N) and day 15 (H). Platelet showed instability on day 9 (L), day 10 (N and H). Mean erythrocyte volume was out of range on day 15 (L), but the N and H control was still stable. The precision and accuracy of Cell Dyn 1400 was in WHO recommended range. We concluded that the precision and accuracy of Cell Dyn 1400 is good. Cell Dyn 16 TL control material was stable until day 9, and its can be recommended to be used as a control material for external quality assessment scheme.

Evaluation of hematological values obtained with reference automated hematology analyzers of six manufacturers.

We evaluated assays of the same fresh blood samples with six different types of reference automated hematology analyzers developed by the following manufacturers: Beckman Coulter, Sysmex, Bayer, Abbott, Nihon Kohden and Horiba. Fresh whole blood samples treated with dipotassium ethylenediaminetetraacetic acid (EDTA K2) were collected from three healthy adult volunteers. The complete blood counts (CBC) including red blood cell count (RBC), hemoglobin (Hgb), hematocrit (Hct), mean corpuscular volume (MCV), white blood cell count (WBC), platelet count (Plt), reticulocyte percentage (Ret) and leukocyte differential counts including % neutrophils (Neu), % lymphocytes (Lym) and % monocytes (Mon) were surveyed with a reference automated hematology analyzer from each manufacturer. The process from sampling to analysis was performed according to procedures in hospital clinical laboratories. RBC, Hgb, Hct and MCV exhibited allowable differences within 5% of mean value among all instruments. Large differences greater than 10% of mean value in WBC, Neu and Lym between Horiba and other manufacturers, and in Plt between Nihon Kohden and other manufacturers, were observed. Ret and Mon exhibited large differences over 10% of mean value among almost all of the instruments tested. This survey suggests that all parameters exhibiting differences greater than 10% of mean value among instruments should be improved for clinical use to ensure good external quality control in blood cell counting and leukocyte differential counting using automated instruments.

Possível interferência da hiperleucocitose no resultado da determinaçäo do hematócrito por aparelho de automaçäo / Possible interference of hiperleucocitosis in results of hematocrit determination from automated analyzers

O uso de aparelhos automatizados na elaboraçäo de hemograma tornou-se uma rotina, porém várias anormalidades hematológicas podem alterar a interpretaçäo do hematócrito, o que implica resultados equivocados. Este trabalho tem como objetivo avaliar o impacto da hiperleucocitose nos resultados oriundos de aparelhos que utilizam método de impedância. A pesquisa foi desenvolvida a partir do equipamento Cobas Argos 5-Diff (Roche) nos hemogramas do Instituto Nacional do Câncer (INCa-RJ). Foram avaliadas 153 amostras, sendo 108 com leucometria abaixo de 100.000 cel/mm elevado ao cubo e 45 acima deste valor. Para cada amostra, foram realizadas cinco leituras no aparelho e cinco micro-hematócritos. As amostras com valor abaixo de 100.000 cel/mm elevado ao cubo näo apresentaram diferença significativa entre as leituras, enquanto as amostras com valores superiores apresentaram diferenças relevantes entreos dois tipos de leitura. A análise estatística realizada permitiu concluir que o aparelho utilizado pode apresentar valores incorretos de hematócrito, no caso de leucometria acima de 100.000 cels/mm elevado ao cubo, sendo, nestes casos, necessária a realizaçäo do micro-hematócrito para confirmaçäo

Utility of HemoCue in estimation of hemoglobin against standard blood cell counter method.

INTRODUCTION: There are various methods which have been recommended for haemoglobin (Hb) estimation for assessment of anaemia. Each method has it's advantages and limitations. OBJECTIVE: The present study was conducted to assess the utility of HemoCue, a recent method, in assessment of haemoglobin against the standard blood cell counter (BCC) method. METHODOLOGY: Eighty five venous blood samples were collected, Hb was assessed in each by utilising the HemoCue and BCC methods. The results were compared utilising the standard statistical methods. RESULTS: The HemoCue provided consistently higher value in comparison to BCC by a margin of 0.5 g/dl. It is known that the correlation coefficient is independent of change of origin but mean gets affected. Therefore, we can subtract 0.5 g/dl from the Hb estimates achieved by HemoCue through which all conditions regarding better agreement between both methods get satisfied. The two assays correlated well. CONCLUSION: The HemoCue method was found more rapid and easy in assessment of total hemoglobin as compared to blood cell counter method with an adjustment of 0.5 per dl.

External quality control program of four blood cell counters

Objetivo. Evaluar un programa de control de calidad interno-externo en cuatro contadores automatizados. Método. Cada una o dos semanas y durante 14 meses, se midieron seis parámetros de medición directa de tres muestras de sangre fresca en cuatro contadores Coulter. La mediana por parámetro del día de trabajo fue utilizada para detectar inexactitudes, y si el programa de control interno del participante confirmó la inexactitud, se calibró el parámetro. Resultados. en 21 de 22 instancias, el programa interno confirmó una inexactitud de un parámetro que se recalibró (cuatro cuentas de leucocitos y cinco de eritrocitos, cinco hemoglobinas, siete volúmenes eritrocíticos). En estos cuatro parámetros sólo hubo pequeñas diferencias entre el contador que menos contó y el que más contó en el análisis global. Contrariamente, los cuatro contadores, con una excepción, difirieron entre sí en los dos parámetros que no pueden ser recalibrados por el usuario (volumen plaquetario, anchura de distribución eritrocítica). Conclusiones. 1. El programa contribuyó a una buena prescisión y exactitud intracontadores y a una buena concordancia intercontadores en los parámetros capaces de ser recalibrados por el ususario. 2. Las diferencias intercontadores en anchura de distribución fueron los suficientemente grande (hasta 9 por ciento) para afectar su interpretación clínica. Esto obliga a establecer valores de referencia de anchura para cada aparato

Control cells for both electrical impedance and light scattering automated hematological analyzers: preparation from normal and thalassemic blood samples.

The study on quality control of automated blood cell analyzers, Technicon H*1 and Coulter MAXM by using three separately self-prepared control cells was extensively investigated. The three parts of control cells are pseudo-leukocyte and fixed platelets, which are fixed by glutaraldehyde, and control red cells from normal and thalassemic patients preserved and anticoagulated in CPD or CPDA-1. The Technicon H*1 system was based on the principle of light scattering but the Coulter MAXM was based on the principle of electrical impedance for cell counting and measurement. The self-prepared control cells can be satisfactorily utilized as control for each system with statistically significant difference (p < 0.05) for both systems. The expired dates for control cells are different in both systems and should be determined for each system specifically. The control red cells prepared from thalassemic patients were quite satisfactorily useful as an abnormal control for both systems during this study.

Control de calidad en la citometría hemática automatizada en el analizador de hematología modelo ®Coulter STKS¼ / Quality control in the automated hematic citometry with the ®Coulter STKS¼ analizer

Todo laboratorio debe contar con un programa que garantice la calidad de sus resultados. Con el objeto de comprobar si los resultados de las citometrías hemáticas procesadas en la Sección de Patología Clínica del Hospital Central Militar son confiable. Se valoró el Control de Calidad Interno (CCI), del analizador hematológico Coulter Modelo STKS, durante un año, utilizando muestras comerciales (Sangre Control 5C), con niveles normales, altos y bajos en cuanto al hemograma completo y diferencial leucocitaria. El coeficiente de variación (CV) obtenido es menor al reportado en la literatura por lo que respecta al hemograma, en cuanto a la diferencial leucocitaria sólo se encontró diferencia significativa en el número de eosinófilos en el nivel normal; y en eosinófilos, linfocitos y neutrófilos en el nivel bajo. Existe correlación al comparar en muestras normales la diferencial leucociatria automatizada y manual en neutrófilos, linfocitos y eosinófilos, pero no en monocitos y basófilos

Thalassemic red cells determined by different technology of blood cell analysers.

Blood samples from 12 normal cases and 30 thalassemic patients were analyzed by four blood cell analysers (the H*1, Baker 9,000RX, Coulter JT, and Argos). These four machines had different technology: light scattering using laser beam (H*1) and electronic aperture impedance (Baker 9,000RX, Coulter JT, and Argos). Analysis of normal blood samples showed comparable red cell parameters between these four machines except the value of RDW. When the thalassemic blood samples were determined using these four types of blood cell counters, many red cell parameters showed significant difference. This indicates that thalassemic red cells which are known as the abnormal cells in terms of their volume, shape, intracellular hemoglobin content, and osmotic fragility, responded differently to the reagents and technology of blood cell analysis. Calibration of the instrument using qualitatively abnormal blood sample, is recommended.