Multicenter evaluation of mycobacteria identification by PCR restriction enzyme analysis in laboratories from Latin America and the Caribbean

Leão, Sylvia Cardoso; Bernardelli, Amelia; Cataldi, Angel; Zumárraga, Martín José; Robledo, Jaime; Realpe, Teresa; Mejía, Gloria Isabel; Telles, María Alice da Silva; Chimara, Érica; Velazco, Maritza; Fernández, Jorge Onrubia; Rodrigues, Pamela Araya; Guerrero, Martha Inírida; León, Clara Inés; Porras, Tania B.; Rastogi, Nalin; Goh, Khye Seng; Suffys, Philip Noël; Rocha, Adalgiza da Silva; Netto, Diogo dos Santos; Ritacco, Viviana; López, Beatriz; Barrera, Lucía; Palomino, Juan Carlos; Martin, Anandi; Portaels, Françoise

doi:10.1016/j.mimet.2004.11.015

Cited by 38 publications

(52 citation statements)

References 9 publications

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“…130 It is a simple and rapid identification method and the turnaround time may be 24-48 hours. 131,132 PRA seems to be an efficient method for the identification of mycobacteria to the species level.…”

Section: High-performance Liquid Chromatography (Hplc)mentioning

confidence: 99%

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Dinnes¹,

Deeks²,

Kunst³

et al. 2007

Health Technol Assess

529

361

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Section: High-performance Liquid Chromatography (Hplc)mentioning

confidence: 99%

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Dinnes¹,

Deeks²,

Kunst³

et al. 2007

Health Technol Assess

529

361

View full text Add to dashboard Cite

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“…Tanto para a detecção de DNA micobacteriano nos sedimentos das amostras processadas, como para a identificação dos isolados de micobactérias, utilizou-se a técnica de PRA (PCR-Restriction Enzyme Analysis) com amplificação do gene hsp65 e análise do polimorfismo dos fragmentos de restrição após digestão, com as enzimas BstEII e HaeIII (Telenti et al, Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio vargas -Manaus/AM 1993; Devallois et al, 1997;Silva et al, 2001;Leão et al, 2005).…”

Section: Procedimentos Molecularesunclassified

Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio Vargas - Manaus/AM

et al. 2009

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RESUMOInvestigou-se por métodos bacteriológicos (cultivo) e moleculares (PCR -Restriction Enzyme Analysis, PRA), a presença de micobactérias ambientais em águas de torneira, soluções e luvas cirúrgicas, utilizadas nas etapas dos procedimentos cirúrgicos executados no centro cirúrgico do Hospital Universitário Getulio Vargas (HUGV), na cidade de Manaus-Amazonas/Brasil. Foram colhidas e analisadas 105 amostras sendo: 24 de águas (colhidas das 2 torneiras existentes no centro cirúrgico), 8 de solução de Povidine e 7 de solução de Clorhexidina, que servem para a higienização das mãos dos cirurgiões; 39 de luvas cirúrgicas (superfícies internas e externas); e 27 de soluções que foram efetivamente utilizadas durante o ato cirúrgico. Por método bacteriológico obteve-se 41 isolados micobacterianos apenas de águas das torneiras. Pelo PRA obteve-se a detecção de DNA micobacteriano somente na amostra de água que forneceu acima de 100 colônias de micobactérias por tubo semeado. Os isolados foram identificados como sendo Mycobacterium celatum perfil 2, M. gordonae perfil 3, M. gordonae perfil 6, M. intracellulare perfil 1, M. lentiflavum perfil 3 e M. mucogenicum perfil 1. O encontro de M. mucogenicum, espécie já incriminada em surtos pós-cirúrgicos, indica que devem ser efetuados procedimentos de limpeza e monitoramento em todos os pontos de distribuição de águas, visando à prevenção de surtos de micobacterioses nosocomiais induzidos pelo uso das águas nas diferentes atividades de manuseio ou higienização dos pacientes submetidos a procedimentos invasivos. PALAVRAS-CHAVE:Micobactérias Ambientais, MNT, água, soluções anti-sépticas, luvas, procedimentos cirúrgicos. Investigation of Environmental Mycobacteria in tap water, surgical gloves and antiseptic solutions used in surgical procedures at the Getúlio Vargas University Hospital, Manaus-AM/BrazilABSTRACT Using bacteriological (culture) and molecular (PCR -Restriction Enzyme Analysis, PRA) methods, we investigated the presence of environmental mycobacteria in tap water, antiseptic solutions and surgical gloves, used in carrying out surgical procedures at the Getúlio Vargas University Hospital Surgical Center, in Manaus -Amazonas/Brazil. Samples (105) were collected and analyzed from: tap water (24 -collected from 3 taps in the surgical center); povidone-iodine solution, PVP-I, (8); Chlorhexidine solution (7), that were used to hygienize surgeons' hands; surgical gloves (39); and solutions that were effectively used during the surgical procedure (27). Using bacteriological method 41 mycobacteria were isolated, all from samples of tap water. Using PRA, mycobacterial DNA was detected only in the sample from water that provided over 100 colonies per inoculated culture tubes. The isolated were identified as Mycobacterium celatum pattern II, M. gordonae pattern III, M. gordonae pattern VI, M. intracellulare pattern I, M. lentiflavum pattern III and M. mucogenicum pattern I. The isolating of M. mucogenicum, a species that had already been incriminated for causing post-surgical outb...

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“…Especie n Subtipo (%) Especie n n (%) reproducibilidad entre laboratorios (21). Se ha observado una buena correlación entre PCRanálisis de restricción y los métodos de identificación fenotípica de aislamientos en Brasil, y además, una buena relación costo-beneficio (17).…”

Section: Identificación Genotípica Identificación Fenotípica Concordaunclassified

“…Ante la facilidad del método de PCR-análisis de restricción, en el mundo se han realizado una serie de estudios para lograr su estandarización y probar la reproducibilidad de la técnica (21). La principal ventaja de PCR-análisis de restricción es que, siendo un método de amplificación, desde la muestra se puede identificar la especie, en corto tiempo y con gran efectividad (18); en la mayoría de los casos, es capaz de diferenciar entre especies de los grupos de micobacterias, como el complejo M. aviumintracellulare de gran importancia clínica, aunque no diferencia los nuevos miembros del complejo, que son M. chimaera y M. colombiense (22)(23).…”

unclassified

Identificación de Haemophilus influenzae, Streptococcus pneumoniae y Neisseria meningitidis por reacción en cadena de la polimerasa

Parra-Barrera¹,

Castañeda²,

Moreno³

2007

biomedica

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Introducción. Las micobacterias no tuberculosas pueden ser saprofitas, patógenas u oportunistas; las enfermedades más comunes producidas por estos microorganismos son las infecciones posquirúrgicas, principalmente por procedimiento estéticos, infecciones asociadas con catéteres, enfermedades cutáneas diseminadas, enfermedades pulmonares y del sistema nervioso central que afectan especialmente a pacientes infectados con el virus de la inmunodeficiencia humana. La identificación fenotípica de las micobacterias no tuberculosas incluye pruebas microbiológicas y bioquímicas, las cuales pueden tomar varias semanas y algunas veces no logran diferenciar entre los miembros de un complejo. Objetivo. El objetivo fue evaluar la metodología de reacción en cadena de la polimerasaanálisis de restricción, como método de identificación genotípica de micobacterias no tuberculosas aisladas de muestras clínicas que pertenecen a la colección del Instituto Nacional de Salud. Materiales y métodos. Se estudiaron 70 aislamientos clínicos de micobacterias no tuberculosas, criopreservados en glicerol al 50% e identificados mediante metodologías fenotípicas. La identificación genotípica se realizó por reacción en cadena de la polimerasa-análisis de restricción y se evaluó la concordancia entre las metodologías. Resultados. Se obtuvo una concordancia del 100% en la identificación de Mycobacterium terrae, M. szulgai, M. avium, M. chelonae y M. scrofulaceum, en las especies M. fortuitum, M. abscessus M. gordonae y M. intracellulare varió de 44% a 89%; no se obtuvo concordancia en la identificación de las especies M. flavescens y M. malmoense. Conclusiones. El análisis de restricción es una alternativa para la identificación de especies de micobacterias no tuberculosas, rápida, económica y segura para la identificación, que permite la diferenciación entre especies de un complejo y la determinación del subtipo de cada especie.Palabras clave: infecciones atípicas por Mycobacterium, reacción en cadena de la polimerasa, técnicas de diagnóstico molecular, infecciones oportunistas, salud pública.

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Multicenter evaluation of mycobacteria identification by PCR restriction enzyme analysis in laboratories from Latin America and the Caribbean

Cited by 38 publications

References 9 publications

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio Vargas - Manaus/AM

Identificación de Haemophilus influenzae, Streptococcus pneumoniae y Neisseria meningitidis por reacción en cadena de la polimerasa

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