Linearidade e carryover do contador hematológico Horiba ® Abx Micros ESV 60 / Linearity and carryover of automated hematology counter Horiba ® Abx Micros ESV 60

O contador automático hematológico ABX Micros ESX 60 (Horiba Medical 2012) é analisador hematológico veterinário multi-espécie que realiza 50 contagens por hora, libera 18 parâmetros sanguíneos, além de fazer representações gráficas (histogramas) para leucócitos, hemácias e plaquetas. O objetivo deste trabalho é avaliar o desempenho do referido aparelho em relação à linearidade e carryover, através de controle comercial e de amostras de sangue provenientes da rotina do Laboratório de Patologia Clínica Veterinária. De acordo com resultados é possível afirmar que o presente aparelho possui um excelente coeficiente de linearidade (r2=0,99) nos parâmetros de leucócitos, eritrócitos e plaquetas em relação às diluições estudadas. Em relação aos carryover houve excelente desempenho do aparelho, contudo, houve valores não conformes nos parâmetros de CHCM e VPM em uma das metodologias realizadas que pode ser justificada pela limitação da fórmula que não considera a características do equipamento.(AU)

The automated hematology counter ABX Micros 60 ESX (Horiba Medical 2012) is veterinary hematology analyzer multi-species that carries 50 counts per hour releases 18 blood parameters, in addition to graphical representations (histograms) for leukocytes, erythrocytes and platelets. The objective of this study is to evaluate the performance of the apparatus with respect to linearity and carryover through commercial control and blood samples from the routine of Veterinary Clinical Pathology Laboratory. According to results we can say that this device has excellent linearity coefficient (r2=0.99) in leukocyte parameters, erythrocytes and platelets during that time dilutions. Regarding the carryover was excellent device performance, however, was not in conformity values ​​in the parameters of MCHC and VPM in one of the methodologies made that can be justified by the limited formula that does not consider the equipment characteristics.(AU)

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.

New Leukocyte Counting Method of Cerebrospinal Fluid: Using Transparent Ruler Tape / 대한진단검사의학회지

BACKGROUND: To enumerate leukocyte count in cerebrospinal fluid (CSF) is important for diagnosing bacterial meningitis. Using automated hematology analyzer for enumeration of leukocyte in CSF is below the sensitivity, so microscopic hemocytometric method is standard method. But this requires sufficient practical experience and has limitation of accuracy and stability. So we developed new manual method and evaluated it. METHODS: We designed new method using transparent ruler tape. We performed correlation, accuracy and precision test by counting leukocyte in diluted EDTA blood with three methods: new method, Neubauer and Nageotte hemocytometry. Twenty two CSF were used for stability test, which determines leukocyte count according to time (within one hour and after 2, 4 and 12 hr), by new method and Neubauer hemocytometry at room temperature. RESULTS: There was no clinical significant difference between three methods in correlation test, whereas Neubauer and Nageotte hemocytometry showed a bias to underestimation relative to the results obtained with new method in case with low leukocyte count. The new method showed the lowest CV and most accurate result. In stability test, leukocyte counts decreased being 44.4%, 72.1% of initial values after 2 hr, 14.8%, 31.1%, after 4 hr and 4.2%, 8.7%, after 12 hr, by Nageotte hemocytometry and new method, respectively. CONCLUSIONS: The new method we devised is simple, easy and applicable to use in a laboratory and offers advantages of improved precision and stability. It may be sufficient for replacing standard methods for leukocyte counting in CSF.

Evaluación del autoanalizador hematológicos Coulter MAX-M® recuento diferencial leucocitario y alarmas morfológicas en pacientes onco-hematológicos / Evaluation of the coulter MAX-M® leukocyte differential counts and morphology alarms in haematology patients

Los recuentos celulares analizados por equipos hematológicos han sido validados desde hace tiempo por sus buenos resultados comparativos. El propósito de este estudio es evaluar el Recuento Leucocitario Diferencial y las alarmas morfológicas entregadas por el autoanalizador hematológico Coulter MAX-M®. Para esto se emplearon 100 muestras provenientes del servicio de Onco-Hematología del Hospital San Juan de Dios (Santiago, Chile) y se compararon con el Método Óptico Convencional. El Recuento Leucocitario Diferencial realizado por el autoanalizador fue comparable con el método manual, con correlaciones muy buenas para Neutrófilos (R² = 0,893), Linfocitos (R² = 0,847) y Eosinófilos (R² = 0,801). Los Monocitos presentaron una buena correlación por ambos métodos (R² = 0,759), pero los Basófilos no tuvieron asociación lineal entre ambos métodos (R² = 0,06). Las alarmas morfológicas definitivas detectadas por el equipo Coulter MAX-M® discreparon al ser comparadas con el método manual, pero las alarmas morfológicas de sospecha coincidieron con las alteraciones encontradas al microscopio. Un 9,8 por ciento de los Hemogramas tenían algún tipo de alteración morfológica, pero el equipo no las detectó, por lo que aunque este equipo es un instrumento confiable, es necesario que una persona calificada y experimentada revise los frontis sanguíneos de todas las muestras por el método manual, con la finalidad de buscar las alteraciones que el equipo no detecta.

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.

Quality control in a manual and an automated leukocyte differential count.

Quality control (QC) has been introduced in laboratories, and QC surveys in leukocyte differential count to enhance quality have been performed by College of American Pathologists, Japanese Association of Medical Technologists, Osaka Medical Association and manufacturers. The results of QC survey in a manual leukocyte differential count indicated problems on the differentiation of segmented neutrophils and band neutrophils and the detection of pathological blood cells on blood smear. While the results of QC survey in an automated leukocyte differential count performed by same manufacturer with an automated blood cell counter were satisfactory, however, there was a difference in leukocyte differential cell counts among laboratories with other manufacturer's instruments because the synthetic blood material used in QC is an exclusive item for an instrument. It is necessary to further reeducate the medical technologists in order to improve morphological performance, and to standardize the synthetic blood material for compatibility with various automated blood cell counters.

Evaluación de la cuenta monocitaria por método automatizado VCS / Evaluation of monocyte count by the automatized

Antecedentes: En un trabajo previo, evaluamos la confiabilidad de la cuenta diferencial automatizada del analizador Coulter Jr con fundamento volumétrico en comparación con el método microscópico manual; encontramos que en casos de pacientes hospitalizados existe una muy buena correlación en granulocitos (r=0.928) y linfocitos (r=0.950), con malos resultados en mononucleares (p < 0.005); fenómeno explicable porque de manera columétrica los monocitos resultan indistinguibles de eosinófilos, basófilos, linfocitos atípicos y células inmaduras lo que conduce a tener que revisar en el microscopio un elevado porcentaje de laminillas. Objetivo: Valorar la confiabilidad de la cuenta de monocitos del método automatizado VCS(V=Volumen, C=Conductancia y S=Dispersión láser) del equipo Coulter MaxM. Tipo de estudio: Prospectivo, comparativo, analítico, en un periodo comprendido entre noviembre de 1993 y enero de 1994. Pacientes y métodos: Se procesaron un promedio de cinco muestras aleatorias diarias, de pacientes hospitalizados y del servicio de urgencias, hasta alcanzar un total de 100 casos. Las muestras de sangre venosa se obtuvieron en tubo Vacutainer K3-EDTA y fueron analizadas en un contador Coulter MaxM y en un Coulter Jr; posteriormente se tiñó un frotis sanguíneo con colorante de Wright y se realizó una cuenta diferencial en 100 células con ocular de 10x y objetivo de 100x. Los resultados obtenidos se correlacionaron estadísticamente. Resultados: En 14 por ciento de las laminillas se encontró una monocitosis de más del 10 por ciento (> 800/mm3). Con los analizadores se encontró una buena correlación en el MaxM (r=0.99) y mala con el Jr (r=0.198). En el Coulter Max se estableció valor predictivo negativo de 99 por ciento, sensibilidad 94 por ciento, especificidad 87 por ciento, con eficiencia de 91 por ciento. Conclusión: Los resultados del Coulter MaxM con tecnología VCS son superiores a los del Coulter Jr, demostrándose así un avance significativo de la tecnología. Aunque estos instrumentos son incapaces de reemplazar totalmente al microscopio, representa una valiosa herramienta para la solución del problema del alto volumen de estudios

Lymphocyte changes in secondary dengue fever: use of the Technicon H*1 to monitor progress of infection.

Secondary dengue fever as dengue hemorrhagic fever or dengue shock syndrome is a potentially fatal complication of an infection that presents with few clinical signs that help in the diagnosis. Previous workers have reported the value of buffy coat determinations of atypical lymphocytes as an aid to the diagnosis. We report here the use of an automated white blood cell differential counter, the Technicon H*1 in the monitoring of the atypical lymphocyte count as a measure of the progress of the infection in a retrospective study of serial full blood counts in 45 serologically confirmed patients. Technicon H*1 'basophil' and large unstained cell counts and manual atypical lymphocyte counts rose in tandem with the drop in platelets and decreased when the platelets recovered. In a sub-study, the atypical lymphocytes were immunophenotyped and found to be predominantly derived from T lymphocytes. We conclude that the Technicon H*1 is a useful monitor of the activity of the infection in dengue hemorrhagic fever.

Contagem específica de células do líqüido céfalo-raquideano em câmara usando coloraçäo supravital / Specific cerebrospinal fluid cells count in counting chamber using supravital staining

Os autores adaptaram ao estudo da citologia do LCR o líquido diluidor de Anguiano e Aneira, proposto para realizaçäo do hemograma em câmara de contagem. O procedimento técnico varia conforme o aspecto do LCR e de acordo com o número global de células. A morfologia das células à técnica de coloraçäo supravital (TCSV) é descirta pormenorizadamente. Utilizando a técnica de Mores-Rêgo e Fernandes os autores determinaram os desvios padröes para cada tipo de células e tabularam o percentual de células determinado na lâmina corada em cada caso. Demonstraram que os valores obtidos pela TCSV caíram dentro dos limites de tolerância calculados sendo, portanto, os valores das contagens equivalentes aos encontrados nas contagens feitas em lâminas, tomadas, estas, como padräo. Com a experiência adquirida durante 5 anos de utilizaçäo da TCSV, os autores ea recomendam como técnica de eleiçäo para a contagem diferencial da célula do LCR