Reticulocyte count: comparison among methods

Introduction: The automated counting of reticulocytes has some advantages over the manual method routinely used in clinical laboratories. Technological innovations provide more statistically reliable results, while optimizing the time to perform this test. However, the cost for implementing the automated procedure in laboratory routines still constitutes a barrier to its use in small- and medium-size Brazilian laboratories. Objective: This study evaluated the performance of a new laboratory protocol for reticulocyte counting by flow cytometry using acridine orange (FC/AO), compared with the manual method and with another automated one by flow cytometry using the commercial kit BD Retic-Count (FC/RC) Conclusion: The results showed that, besides being comparable to the manual method, still considered standard, the evaluated new protocol is economically more advantageous than the automated methods currently available, and its cost is comparable to that of the manual method for laboratories that already have appropriate equipment and infrastructure

Evaluation of the interference of platelets in reticulocyte counting by flow cytometry using thiazole orange

ABSTRACT Introduction: The reticulocyte count by flow cytometry (FC) - an automated counting method - can present errors due to the presence of interfering factors, contributing to a slight increase in results. However, automated methods have large advantages over the manual method, taken as reference, what justifies efforts to improve their quality. Objective: Evaluate platelet interference with the reticulocyte count by FC, using thiazole orange (TO) (FC/TO). Materials and methods: The method of reticulocyte count by FC/TO and a modified automated equivalent method, which excluded CD61-positive cells (platelets) from analysis (FC/TO/MOD), were compared to the manual method. Conclusion: Results were analyzed according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI) to assess interchangeability between the methods, by linear regression analysis and paired t-test. The exclusion of interfering fragments from result analysis by the modified method produced results in closer proximity to those of the reference method

Flow cytometry reticulocyte counting using acridine orange: validation of a new protocol

Introduction: Currently, the reticulocyte counting is a challenge for clinical laboratories in Brazil, mainly for the ordinary ones, which still use the manual method. This method has some limitations, since it consists of a laborious method, time consuming, with low accuracy. Objectives: This study has developed and evaluated the performance of a New Laboratory Protocol for flow cytometry (FC) reticulocytes counting using acridine orange (AO) as dye, aiming to standardize a more precise, easy, fast implementation, and low cost protocol. After standardization of the New Protocol (FC/AO), it was compared with the manual method. The results were analyzed according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS), now known as Clinical and Laboratory Standards Institute (CLSI), to evaluate the interchangeability of methods in linear regression analysis and paired t test, besides other quality control tests. Conclusion: Based on these results concerning to the correlation between the methods and the tests related to quality control, we can admit that FC/AO for reticulocyte counting shows undeniable advantages when compared to the preexisting manual method

Flow cytometry reticulocyte counting using acridine orange: validation of a new protocol

Submitted by Nuzia Santos ([email protected]) on 2015-03-26T13:19:34Z No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T13:19:49Z (GMT) No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T13:26:11Z (GMT) No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Made available in DSpace on 2015-03-26T13:26:11Z (GMT). No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5) Previous issue date: 2014Universidade Federal de Minas Gerais. Faculdade de Farmácia. Belo Horizonte, MG, Brasil/Prefeitura de Belo Horizonte. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clinicas e Toxicologicas. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clinicas e Toxicologicas. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Belo Horizonte, MG, BrasilCentro Universitário Newton Paiva. Belo Horizonte, MG, BrasilLaboratório de Patologia Clínica São Paulo. Serviço de Citometria. Belo Horizonte, MG, BrasilFundação Hemominas. Hematologia. Belo Horizonte, MG, BrasilLaboratório de Patologia Clínica São Paulo. Hematologia Clinica. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilIntrodução: Atualmente, a contagem de reticulócitos representa um desafio para os laboratórios clínicos no Brasil, principalmente os de pequeno e médio porte, nos quais ainda se utiliza o método manual. Este método apresenta algumas limitações, classificando-se como tedioso, demorado e de baixa precisão. Objetivos: O presente estudo desenvolveu e avaliou o desempenho de um novo protocolo laboratorial para contagem de reticulócitos por citometria de fluxo (CF) utilizando acridine orange (AO) como corante, visando padronizar um protocolo preciso, de fácil e rápida execução e custo acessível. Após a padronização do novo protocolo desenvolvido (CF/AO), fez-se a comparação com o método manual. Os resultados foram analisados de acordo com recomendações do National Committee for Clinical Laboratory Standards (NCCLS), atualmente Clinical and Laboratory Standards Institute (CLSI), para avaliar a intercambialidade entre os métodos, por meio da análise de regressão linear e teste t pareado, além de outros testes de controle de qualidade. Conclusão: Diante dos resultados obtidos referentes à correlação entre os métodos e os testes voltados ao controle de qualidade, pode-se admitir que o CF/AO estabelecido para contagem de reticulócitos possui vantagens inegáveis quando comparado com o método manual.Introduction: Currently, the reticulocyte counting is a challenge for clinical laboratories in Brazil, mainly for the ordinary ones, which still use the manual method. This method has some limitations, since it consists of a laborious method, time consuming, with low accuracy. Objectives: This study has developed and evaluated the performance of a New Laboratory Protocol for flow cytometry (FC) reticulocytes counting using acridine orange (AO) as dye, aiming to standardize a more precise, easy, fast implementation, and low cost protocol. After standardization of the New Protocol (FC/AO), it was compared with the manual method. The results were analyzed according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS), now known as Clinical and Laboratory Standards Institute (CLSI), to evaluate the interchangeability of methods in linear regression analysis and paired t test, besides other quality control tests. Conclusion: Based on these results concerning to the correlation between the methods and the tests related to quality control, we can admit that FC/AO for reticulocyte counting shows undeniable advantages when compared to the preexisting manual method