Método fenotípico automatizado versus método manual na identificação de microorganismos isolados de hemoculturas: desfechos clínicos e microbiológicos / Automated phenotypic method versus manual method in the identification of microorganisms isolated from blood cultures: clinical and microbiological outcomes

INTRODUÇÃO: A automação laboratorial é cada vez mais utilizada em microbiologia, no entanto, poucos estudos avaliam desfechos clínicos em comparação aos métodos tradicionais. No Brasil, nenhum estudo com esse objetivo foi detectado. OBJETIVO: Analisar os impactos clínicos e microbiológicos após implantação de método fenotípico automatizado em um serviço de microbiologia. MÉTODOS: Realizamos estudo observacional e retrospectivo no laboratório de microbiologia referente a exame de hemocultura de pacientes da Unidade de Terapia Intensiva (UTI). Os dados foram coletados de pacientes internados entre janeiro/2014 a dezembro/2015. Analisou-se o tempo de internação, número de terapias empíricas, óbitos e dados relacionados ao isolamento microbiológico. A amostra foi obtida por conveniência. Para a comparação entre os desfechos foram empregados os testes t de Student e Qui-quadrado de Pearson. O programa empregado foi o Stata release, versão 11, sendo considerados significativos valores de p<0,05. RESULTADOS: Foram avaliados 472 pacientes. Não houve redução na prescrição empírica de antimicrobianos (54,7% vs 45,3%; p=0,33), tempo de internação na UTI (14,5 dias vs 15,8 dias p=0,78) e na taxa de óbitos (54,4% vs 45,6%; p=0,36). Similarmente, o perfil de agentes isolados em ambos os métodos não parece ser discrepante, no entanto, houve um aumento de 44,7% no número de isolados microbianos (76 vs 110) com melhor caracterização dos mesmos. CONCLUSÃO: A automação do laboratório de microbiologia não impactou no tempo de internação, mortalidade na UTI e no número de terapias empíricas. No entanto, a identificação e o isolamento de microrganismos melhoraram.

INTRODUCTION: Automation is increasingly used in microbiology laboratory, however, few studies assessed clinical outcomes compared to traditional methods. In Brazil, no studies with this objective were detected. OBJECTIVE: To analyze the clinical and microbiological impacts after implantation of an automated phenotypic method in a microbiology service. METHODS: Observational and retrospective study carried out on the microbiology laboratory involving blood culture test from intensive care unit (ICU) patients. Data were collected from hospitalized patients between January 2014 and December 2015. The length of hospitalization, number of empirical therapies, deaths and information related to microbiological isolation were analyzed. The sample was obtained by convenience. Pearson's Chisquare and Student's t-tests were used to compare outcomes. The program used was the Stata release, version 11, being considered significant values of p<0.05. RESULTS: A total of 472 patients were evaluated. There was no reduction in the empirical prescription of antimicrobials (54.7% vs 45.3%; p=0.33), ICU stay (14.5 days vs 15.8 days; p=0.78) and mortality (54.4% vs 45.6%; p=0.36). Similarly, profile of isolated agents in both methods did not appear to be discrepant, however, there was an increase of 44.7% in the number of microbial isolates (76 vs 110) and a better characterization of them. CONCLUSION: The microbiology laboratory automation did not modify the length of stay, ICU mortality and the number of empirical therapies. However, identification and isolation of microorganisms was improved.

Increase in the diagnosis of mycobacteremia after the implementation of semi-automated blood culture at a Brazilian public health laboratory

Abstract INTRODUCTION The prevalence of hematogenous dissemination of mycobacteria is high in immunosuppressed patients. The isolation of mycobacteria in culture remains the standard procedure. METHODS This is a cross-sectional study based on the results of solid (Löwenstein-Jensen medium) and semi-automated liquid (BACTEC 9240) blood cultures, obtained from the Lacen-GO database. RESULTS The implementation of a semi-automated procedure resulted in an increase of 61.5% and 350.0% in the positive results for Mycobacterium tuberculosis complex and nontuberculous mycobacteria, respectively. This technique also accelerated the detection of positive results. CONCLUSIONS Semi-automated liquid blood culture showed a better performance in the diagnosis of mycobacteremia.

Experiencia en el despliegue del piloto de galen clínicas en los laboratorios del instituto de hematología e inmunología / Experience in deploying of pilot galen clínicas laboratories in the institute of hematology and immunology

SOFTEL, es la empresa encargada por el Ministerio de las Comunicaciones de informatizar las entidades de Salud Pública. Para ello se han desarrollado varias aplicaciones, para ser desplegadas en hospitales, clínicas, policlínicos y bancos de sangre, fundamentalmente. Específicamente los laboratorios constituyen un área muy sensible debido a que en ellos se realizan todos los medios diagnósticos, que resultan de gran utilidad el almacenamiento de esta información para futuros estudios de los pacientes. Este trabajo expone la experiencia obtenida en el despliegue de dicho módulo, como una primera experiencia, conocida como despliegue de piloto o prueba beta. Esta experiencia sirve de base para las futuras instalaciones en otros clientes, reportó una serie de inconformidades que se resolvieron antes de concluir el mismo y finalmente quedo concluido el despliegue. Este tuvo complicaciones relacionadas con la existencia de laboratorios fuera del área de cobertura del Instituto y la preparación de las interfaces con los diferentes auto analizadores(AU)

SOFTEL is the enterprise commissioned by the Ministry of Communications to computerize public health institutions. For that has been developed several applications to be deployed in hospitals, clinics and blood banks, basically. Specifically, laboratories are a very sensitive area because in them all the diagnostic tools are used, so it is useful storage of this information for future studies of patients. This paper describes the experience gained in the deployment of the module, as a first experience, known as pilot or deployment of beta testing. This experience, which forms the basis for future installations on other customers, reported a series of disagrees which were resolved before the end of the same and finally the deployment was completed. This had complications related to the existence of laboratories outside the coverage area of the Institute and the preparation of the interfaces with the different analyzers(AU)

El conteo automático de reticulocitos: una herramienta de uso diagnóstico, clínico e investigativo / Automated reticulocyte count: a tool for diagnostic, clinical and research use

Se revisan los antecedentes históricos del conteo de reticulocitos como determinación indispensable en el laboratorio de hematología para la evaluación de la actividad eritropoyética durante la clasificación, diagnóstico y monitoreo de la respuesta terapéutica en distintos trastornos y situaciones clínicas, principalmente en casos de anemias. Se describe el tránsito del tradicional método de conteo de reticulocitos manual al método automatizado y la integración de los parámetros reticulocitarios al hemograma automatizado actual; además, se analizan las desventajas del método de recuento manual y las ventajas del método automatizado, así como los principios de detección en que se basa el conteo electrónico de reticulocitos. Con relación a los parámetros reticulocitarios, se describe su medición, cálculo y unidades de medida; también se resalta la importancia de la fracción de reticulocitos inmaduros y del contenido de hemoglobina reticulocitaria como variables de mayor uso clínico e investigativo en la evaluación de la respuesta medular ante diversos trastornos clínicos y protocolos terapéuticos. Por último, se alude a la necesidad del conocimiento y empleo de las variables reticulocitarias en la práctica clínica de rutina por parte de los clínicos y especialistas en hematología.

The historical background of reticulocyte count is reviewed as an essential determination in the laboratory of hematology for the evaluation of erythropoietic activity during classification, diagnosis and monitoring of therapeutic response of different conditions and clinical situations are also reviewed, especially in anemia The transition from traditional manual reticulocyte counting method to automated method and integration of the reticulocyte parameters to current automated complete blood count are described. The disadvantages of manual method and the advantages of automated methods are cited, as well as detection principles in which electronic reticulocyte count is based. Regarding reticulocyte parameters, measurement, calculation and units are described. The importance of immature reticulocyte fraction and reticulocyte hemoglobin content as variables most clinical and research use in evaluating bone marrow response to various clinical disorders and therapeutic protocols are highlighted. Finally, the need for knowledge and use of reticulocyte variables in routine clinical practice by clinicians and hematologist is referred.

Evaluación de la incorporación de un sistema automatizado de cultivo microbiológico en un hospital público de Nuquén / Assessment of the incorporation of an automated microbiological culture system in a public hospital of Neuquén

En este artículo se presenta un informe rápido de evaluación de tecnología sanitaria sobre la incorporación de un sistema de cultivo microbiano automatizado para la realización de pruebas de identificación y sensibilidad antibiótica de gérmenes en el laboratorio de microbiología de un hospital público de alta complejidad de la provincia del Neuquén.(AU)

Impact of Implementation of an Automated Liquid Culture System on Diagnosis of Tuberculous Pleurisy

This study was conducted to evaluate the impact of implementation of an automated liquid culture system on the diagnosis of tuberculous pleurisy in an HIV-uninfected patient population. We retrospectively compared the culture yield, time to positivity, and contamination rate of pleural effusion samples in the BACTEC Mycobacteria Growth Indicator Tube 960 (MGIT) and Ogawa media among patients with tuberculous pleurisy. Out of 104 effusion samples, 43 (41.3%) were culture positive on either the MGIT or the Ogawa media. The culture yield of MGIT was higher (40.4%, 42/104) than that of Ogawa media (18.3%, 19/104) (P<0.001). One of the samples was positive only on the Ogawa medium. The median time to positivity was faster in the MGIT (18 days, range 8-32 days) than in the Ogawa media (37 days, range 20-59 days) (P<0.001). No contamination or growth of nontuberculous mycobacterium was observed on either of the culture media. In conclusion, the automated liquid culture system could provide approximately twice as high yields and fast results in effusion culture, compared to solid media. Supplemental solid media may have a limited impact on maximizing sensitivity in effusion culture; however, further studies are required.

Letter to the editor: Respective Contribution of Liquid and Solid Media to Mycobacterial Yields from Pleural Fluid in Tuberculous Pleural Effusion

No abstract available.

Evaluación de un sistema automatizado de siembra de orinas para urocultivos / Evaluation of an automated streaking system of urine samples for urine cultures

Introduction: Automated systems have simplified laboratory workflow, improved standardization, traceability and diminished human errors and workload. Although microbiology laboratories have little automation, in recent years new tools for automating pre analytical steps have appeared. Objectives: To assess the performance of an automated streaking machine for urine cultures and its agreement with the conventional manual plating method for semi quantitative colony counts. Materials and Methods: 495 urine samples for urinary culture were inoculated in CPS® agar using our standard protocol and the PREVI™ Isola. Rates of positivity, negativity, polymicrobial growth, bacterial species, colony counts and re-isolation requirements were compared. Results: Agreement was achieved in 98.97% of the positive/negative results, in 99.39% of the polymicrobial growth, 99.76% of bacterial species isolated and in 98.56 % of colony counts. The need for re-isolation of colonies decreased from 12.1% to 1.1% using the automated system. Discussion: PREVI™ Isola's performance was as expected, time saving and improving bacterial isolation. It represents a helpful tool for laboratory automation.

Introducción: Los sistemas automatizados han facilitado el flujo de trabajo, mejorado la estandarización, la trazabilidad, disminuido el error humano y la carga de trabajo en los laboratorios. A pesar de que la microbiología ha permanecido poco automatizada, en los últimos años han aparecido nuevas herramientas para la automatización de la etapa pre analítica. Objetivos: Evaluar el desempeño de un sistema automatizado de siembra de urocultivos y la concordancia con la siembra manual convencional en el recuento semicuantitativo de colonias. Materiales y Métodos: 495 muestras de orinas fueron sembradas según nuestro protocolo habitual y comparadas con las placas de CPS® obtenidas con PREVI™ Isola en cuanto a positividad/negatividad, muestras polimicrobianas, especies de bacterias aisladas, recuentos y necesidad de resembrar. Resultados: Hubo concordancia en 98,97% de los positivos y negativos, en 99,39% de las muestras polimicrobianas, en 99,76% de las especies aisladas y en 98,56% de los recuentos. La necesidad de resiembra disminuyo de 12,1% a un 1,1% usando este sistema automatizado. Discusión: El desempeño de PREVI™ Isola fue el esperado, mejorando el aislamiento bacteriano y el tiempo requerido y representa una buena herramienta para la automatización de laboratorios.

Avaliação do desempenho analítico do analisador hematológico Sysmex® XE-2100D / Performance evaluation of the Sysmex XE-2100D automated hematology analyzer

Introduction: The Sysmex® XE-2100D is a multiparameter hematology analyzer designed for hematology testing in samples with ethylenediamine tetraacetic acid (EDTA). Objectives: Considering the importance of this hematology analyzer for clinical and laboratory practice, the objective of this study was to evaluate its analytical performance, comparing the obtained results with quality specifications described in literature. Material and method: In the evaluation of analytical performance, according to recommendations of the document H26-A2 of the Clinical and Laboratory Standards Institute (CLSI), intra-run imprecision, inter-run imprecision, linearity, carryover, autosampler evaluation, clinical sensitivity of the atypical lymphocytes flag (n = 400 samples) were included, as well as the comparison between automated and manual leukocyte differential count (n = 400 samples), based on an adaptation of the document H20-A2 of CLSI. Results: Repeatability, reproducibility, linearity and carryover were satisfactory according to the manufacturer's specifications. The clinical sensitivity of the atypical lymphocytes flag showed efficiency, sensitivity and specificity of 92.5%, 65.2% and 94.1% respectively. The correlation coefficients between the automated and manual differential counts of neutrophils, lymphocytes, monocytes, eosinophils and basophils were 0.991, 0.99, 0.872, 0.974 and 0.557, respectively. Conclusions: The results were in accordance with quality specifications described in literature, indicating reliability in Sysmex® XE-2100D. This fact ensures certainty to both laboratory professionals and medical staff. We conclude that the Sysmex® XE-2100D showed excellent analytical performance, and is useful to provide reliable hematology data...

Introdução: O Sysmex® XE-2100D é um analisador hematológico multiparamétrico destinado à realização de testes hematológicos em sangue anticoagulado com ácido etilenodiamino tetra-acético (EDTA). Objetivos: Considerando a sua importância na prática clínica e laboratorial, o objetivo deste estudo foi avaliar seu desempenho analítico, comparando os resultados obtidos com especificações de qualidade descritas na literatura. Material e método: Na avaliação de desempenho analítico, conforme recomendações do documento H26-A2 do Clinical and Laboratory Standards Institute (CLSI), foram incluídos ensaios de verificação da imprecisão intraensaio ou repetitividade, imprecisão entre ensaios ou reprodutibilidade, linearidade, carryover (arraste), avaliação do mecanismo homogeneizador de amostras, sensibilidade clínica do alerta morfológico (flag) de linfócitos atípicos (n = 400 amostras) e a comparação entre a contagem diferencial de leucócitos automatizada e a manual (n = 400 amostras), baseada em uma adaptação do documento H20-A2 do CLSI. Resultados: Os ensaios de verificação da repetitividade, reprodutibilidade, linearidade, carryover (arraste) foram satisfatórios conforme especificações do fabricante. O ensaio de sensibilidade clínica do alerta morfológico (flag) de linfócitos atípicos mostrou eficiência, sensibilidade e especificidade de 92,5%; 65,2% e 94,1% respectivamente. Os coeficientes de correlação entre as contagens diferenciais automatizadas e manuais de neutrófilos, linfócitos, monócitos, eosinófilos e basófilos foram de 0,991; 0,99; 0,872; 0,974 e 0,557 respectivamente...

Comparison of the efficiency between two sampling plans for aflatoxins analysis in maize

Variance and performance of two sampling plans for aflatoxins quantification in maize were evaluated. Eight lots of maize were sampled using two plans: manual, using sampling spear for kernels; and automatic, using a continuous flow to collect milled maize. Total variance and sampling, preparation, and analysis variance were determined and compared between plans through multifactor analysis of variance. Four theoretical distribution models were used to compare aflatoxins quantification distributions in eight maize lots. The acceptance and rejection probabilities for a lot under certain aflatoxin concentration were determined using variance and the information on the selected distribution model to build the operational characteristic curves (OC). Sampling and total variance were lower at the automatic plan. The OC curve from the automatic plan reduced both consumer and producer risks in comparison to the manual plan. The automatic plan is more efficient than the manual one because it expresses more accurately the real aflatoxin contamination in maize.

Avances y aplicación clínica de la citometría hemática automatizada / Advances and clinical application of automated hematic cytometry

Se revisan los antecedentes históricos del surgimiento de la automatización en la rama hematológica y los principios de detección utilizados por los contadores automáticos de células sanguíneas, con referencia a su acción combinada en la caracterización de las distintas poblaciones celulares hemáticas. Se describen los componentes básicos de los analizadores hematológicos. Se detalla la obtención, intervalo de referencia y utilidad clínica de las variables de uso común ofrecidas por estos equipos en el estudio de los eritrocitos y trombocitos, referidos a los parámetros más actualizados en el estudio de estas series celulares. En cuanto a los leucocitos, se mencionan los principales parámetros relacionados con el estudio automatizado de uso rutinario, la utilidad de los novedosos principios inmunológicos de detección en la caracterización de esta población celular, así como los logros y obstáculos manifiestos durante el conteo y estudio de las subpoblaciones leucocitarias inmaduras o anómalas. Por último, se exponen las ventajas del uso de los contadores hematológicos para el trabajo actual del laboratorio de hematología y la necesidad del conocimiento de sus potencialidades diagnósticas por parte de los médicos, especialmente los hematólogos e inmunólogos

Historical background on the development of automation in hematology as well as the major detection principles used by automated blood cell counters are reviewed, referring to their combined action in the characterization of various hematic cell populations. The basic components of hematological analyzers are described. Obtention reference interval and clinical utility of commonly used variables provided by these equipments for the study of erythrocytes and thrombocytes are described referring to the most updated parameters for the study of these cell series. The main parameters of the everyday leukocyte automated study, the novel immunological detection principles used in its characterization, as well as the achievements and obstacles found during this process are mentioned. Finally, the advantages of blood cell counters for the current hematology laboratory work and the need of knowledge of its diagnostic potential by physicians, particularly hematologists and immunologists, are presented

Rapid culture diagnosis of tuberculous lymphadenitis from a tertiary care centre in an endemic nation: Potential and pitfalls.

In spite of low sensitivity and specificity, standard diagnostic algorithm recommends fine needle aspiration cytology (FNAC) and direct microscopic screening for acid-fast bacilli (AFB) for the routine diagnosis of tuberculous lymphadenopathy (LNTB). In this study, the diagnostic utility of liquid broth based automated culture (BacT/ALERT 3D) technique was assessed in comparison with conventional techniques in 89 clinically suspected tubercular lymphadenitis patients. 60% (n = 53) were positive by FNAC and 38.4% (n = 34) demonstrated AFB in smear examination. BacT/ALERT yielded isolation in 43.1% (n = 38) aspirates, confirming tubercular aetiology. We also found six paediatric culture-positive cases which showed negative outcome by both FNAC and smear. Thus, we conclude that culture by BacT/ALERT, may be used for faster yield of Mycobacteria in LNTB, especially in children. Additionally, this could also be used as a platform for further differentiation of Mycobacterium tuberculosis from non-tuberculous mycobacteria (NTM) infection and for testing of anti-tubercular chemotherapeutic agents whenever drug resistance is suspected.

Evaluation of the automated system Vitek2 for identification and antimicrobial susceptibility testing of Brazilian Gram-positive cocci strains

Automated instruments offer many advantages for clinical laboratories. Nevertheless, they can have problems identifying and determining susceptibilities of some pathogens. Vitek® 2 (bioMérieux) is an automated system that was recently introduced to Brazil. We evaluated the performance of this equipment for Brazilian isolates that had been characterized using reference identification and antimicrobial susceptibility testing methods. Ninety-nine strains of Gram-positive cocci from a local reference center collection were analyzed, consisting of 50 coagulasenegative Staphylococcus (CoNS) and 49 Enterococcus and related species. Vitek® 2 correctly identified 79.8 percent (79/99) of the isolates. Oxacillin resistance was detected in 76 percent (19/25) of resistant S. epidermidis strains and in 88 percent (22/25) of other resistant CoNS species strains. Vancomycin resistance was detected in 100 percent (20/20) of resistant Enterococcus and related species strains. Vitek® 2 performed very well for the identification of S. epidermidis and non-epidermidis staphylococci, and for the detection of vancomycin resistance in Enterococcus and related species. However, the system needs improvement in order to provide reliable results for the characterization of some CoNS species, identification of Enterococcus and related species and for detecting oxacillin resistance in CoNS.