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CC BY-NC-ND 4.0 · Rev Bras Ortop (Sao Paulo) 2022; 57(04): 689-696
DOI: 10.1055/s-0041-1732386
Artigo Original
Infectologia

Detecção de microrganismos em dispositivos ortopédicos sonicados clínicos usando cultura convencional e qPCR

Article in several languages: português | English
Victoria Stadler Tasca Ribeiro 
1   Laboratório de Doenças Infecciosas Emergentes (LEID), Escola de Medicina, Departamento de Ciências da Saúde, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil
,
Juliette Cieslinski 
1   Laboratório de Doenças Infecciosas Emergentes (LEID), Escola de Medicina, Departamento de Ciências da Saúde, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil
,
Julia Bertol 
2   School of Medicine, Department of Health Sciences, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil
,
Ana Laura Schumacher 
2   School of Medicine, Department of Health Sciences, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil
,
João Paulo Telles 
2   School of Medicine, Department of Health Sciences, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil
,
Felipe Francisco Tuon 
1   Laboratório de Doenças Infecciosas Emergentes (LEID), Escola de Medicina, Departamento de Ciências da Saúde, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil
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Resumo

Objetivo Avaliar a sensibilidade e a especificidade da reação em cadeia de polimerase em tempo real quantitativa (quantitative real-time polymerase chain reaction, qPCR, em inglês) para a triagem do gene rDNA 16S, com a utilização do fluido sonicado de implantes ortopédicos.

Métodos Um estudo retrospectivo foi realizado em 73 fluidos sonicados obtidos de pacientes com infecção associada aos implantes ortopédicos. As amostras foram submetidas a cultura convencional e a teste molecular utilizando ionização e dessorção a laser assistida por matriz com espectrometria de massa por tempo de voo (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF MS, em inglês) e qPCR para o gene rDNA 16S. Os valores limiares do ciclo foram usados para definir um ponto de corte para a qPCR do gene rDNA 16S para culturas negativas e positivas.

Resultados Não foram observadas diferenças estatísticas entre os grupos de cultura positiva e negativa com base no tempo desde a primeira cirurgia até a infecção (p = 0,958), na idade (p = 0,269), ou nas comorbidades em geral. No entanto, uma diferença estatística foi encontrada entre a duração média do uso de antibióticos antes da remoção do dispositivo (3,41 versus 0,94; p = 0,016). O DNA bacteriano foi identificado em todas as amostras dos fluidos sonicados. Os limiares do ciclo médio de culturas positivas e negativas foram de 25,6 e 27,3, respectivamente (p < 0,001). Como uma ferramenta de diagnóstico, um corte do limite do ciclo de 26,89 demonstrou uma área sob a curva da característica de operação do receptor de 0,877 (p ≤ 0,001).

Conclusão A presença de agentes antimicrobianos por mais de 72 horas diminuiu a positividade da cultura, mas não influenciou os resultados da qPCR. Apesar disso, a amplificação do rDNA 16S pode sobrestimar o diagnóstico de infecção.

Palavras-chave

sonicação - infecções - qPCR - espectrometria de massas por ionização e dessorção a laser assistida por matriz - próteses e implantes

Suporte Financeiro

Este estudo foi apoiado por uma bolsa do MCTIC/CNPq, número 28/2018, nível B.




Publication History

Received: 11 September 2020

Accepted: 19 February 2021

Article published online:
01 October 2021

© 2021. Sociedade Brasileira de Ortopedia e Traumatologia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commecial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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