Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista Brasileira de Ortopedia (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-36162022000400689 |
Resumo: | 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. |
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Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCRsonicationinfectionsqPCRspectrometrymassmatrix-sssisted laser desorption-ionizationprostheses and implantsAbstract 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.Sociedade Brasileira de Ortopedia e Traumatologia2022-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-36162022000400689Revista Brasileira de Ortopedia v.57 n.4 2022reponame:Revista Brasileira de Ortopedia (Online)instname:Sociedade Brasileira de Ortopedia e Traumatologia (SBOT)instacron:SBOT10.1055/s-0041-1732386info:eu-repo/semantics/openAccessRibeiro,Victoria Stadler TascaCieslinski,JulietteBertol,JuliaSchumacher,Ana LauraTelles,João PauloTuon,Felipe Franciscoeng2022-08-30T00:00:00Zoai:scielo:S0102-36162022000400689Revistahttp://www.rbo.org.br/https://old.scielo.br/oai/scielo-oai.php||rbo@sbot.org.br1982-43780102-3616opendoar:2022-08-30T00:00Revista Brasileira de Ortopedia (Online) - Sociedade Brasileira de Ortopedia e Traumatologia (SBOT)false |
dc.title.none.fl_str_mv |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR |
title |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR |
spellingShingle |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR Ribeiro,Victoria Stadler Tasca sonication infections qPCR spectrometry mass matrix-sssisted laser desorption-ionization prostheses and implants |
title_short |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR |
title_full |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR |
title_fullStr |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR |
title_full_unstemmed |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR |
title_sort |
Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR |
author |
Ribeiro,Victoria Stadler Tasca |
author_facet |
Ribeiro,Victoria Stadler Tasca Cieslinski,Juliette Bertol,Julia Schumacher,Ana Laura Telles,João Paulo Tuon,Felipe Francisco |
author_role |
author |
author2 |
Cieslinski,Juliette Bertol,Julia Schumacher,Ana Laura Telles,João Paulo Tuon,Felipe Francisco |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Ribeiro,Victoria Stadler Tasca Cieslinski,Juliette Bertol,Julia Schumacher,Ana Laura Telles,João Paulo Tuon,Felipe Francisco |
dc.subject.por.fl_str_mv |
sonication infections qPCR spectrometry mass matrix-sssisted laser desorption-ionization prostheses and implants |
topic |
sonication infections qPCR spectrometry mass matrix-sssisted laser desorption-ionization prostheses and implants |
description |
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. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-08-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-36162022000400689 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-36162022000400689 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1055/s-0041-1732386 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Ortopedia e Traumatologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Ortopedia e Traumatologia |
dc.source.none.fl_str_mv |
Revista Brasileira de Ortopedia v.57 n.4 2022 reponame:Revista Brasileira de Ortopedia (Online) instname:Sociedade Brasileira de Ortopedia e Traumatologia (SBOT) instacron:SBOT |
instname_str |
Sociedade Brasileira de Ortopedia e Traumatologia (SBOT) |
instacron_str |
SBOT |
institution |
SBOT |
reponame_str |
Revista Brasileira de Ortopedia (Online) |
collection |
Revista Brasileira de Ortopedia (Online) |
repository.name.fl_str_mv |
Revista Brasileira de Ortopedia (Online) - Sociedade Brasileira de Ortopedia e Traumatologia (SBOT) |
repository.mail.fl_str_mv |
||rbo@sbot.org.br |
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1752122363604893696 |