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

Victoria Stadler Tasca Ribeiro; Juliette Cieslinski; Julia Bertol; Ana Laura Schumacher; João Paulo Telles; Felipe Francisco Tuon

doi:10.1055/s-0041-1732386

Revista Brasileira de Ortopedia, Inhaltsverzeichnis

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

Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR

Artikel in mehreren Sprachen: português | English

Victoria Stadler Tasca Ribeiro

¹Laboratório de Doenças Infecciosas Emergentes (LEID), Escola de Medicina, Departamento de Ciências da Saúde, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil

,

Juliette Cieslinski

¹Laboratório de Doenças Infecciosas Emergentes (LEID), Escola de Medicina, Departamento de Ciências da Saúde, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil

,

Julia Bertol

²School of Medicine, Department of Health Sciences, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil

,

Ana Laura Schumacher

²School of Medicine, Department of Health Sciences, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil

,

João Paulo Telles

²School of Medicine, Department of Health Sciences, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil

,

Felipe Francisco Tuon

¹Laboratório de Doenças Infecciosas Emergentes (LEID), Escola de Medicina, Departamento de Ciências da Saúde, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, PR, Brasil

› Institutsangaben

Abstract

Objective To evaluate the sensitivity and specificity of the quantitative real-time polymerase chain reaction (qPCR) for 16S rDNA gene screening using sonicated fluid from orthopedic implants.

Methods A retrospective study was conducted on 73 sonicated fluids obtained from patients with infection associated with orthopedic implants. The samples were subjected to conventional culture and molecular testing using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and qPCR for 16S rDNA. The cycle threshold values were used to define a cut-off of the qPCR of the 16S rDNA for negative and positive cultures.

Results No statistical differences were observed between the positive and negative culture groups based on the time from the first surgery to infection (p = 0.958), age (p = 0.269), or general comorbidities. Nevertheless, a statistical difference was found between the mean duration of antibiotic use before device removal (3.41 versus 0.94; p = 0.016). Bacterial DNA was identified in every sample from the sonicated fluids. The median cycle thresholds of the positive and negative cultures were of 25.6 and 27.3 respectively (p < 0.001). As a diagnostic tool, a cycle threshold cut-off of 26.89 demonstrated an area under the curve of the receiver operating characteristic of 0.877 (p ≤ 0.001).

Conclusion The presence of antimicrobial agents for more than 72 hours decreased culture positivity, but did not influence the qPCR results. Despite this, amplification of the 16S rDNA may overestimate infection diagnosis.

Keywords

sonication - infections - qPCR - spectrometry, mass, matrix-sssisted laser desorption-ionization - prostheses and implants

Volltext

Referenzen

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