Inoculation of sonicate fluid into blood culture bottles improves microbial detection in patients with orthopedic bone and soft tissue infections of the upper and lower extremities

Toyama, Takeyasu; Ueda, Narumi; Hamada, Yoshitaka; Okuda, Kuniaki; Saito, Takanori

doi:10.1016/j.jos.2021.11.012

Cited by 2 publications

(3 citation statements)

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“…The device was lled with tap water containing a special surfactant (Tickopur TR 3; DR. H. STAMM GmbH, Berlin, Germany) for 15 minutes before sonication. Each sample bottle containing infectious tissues was vortexed for 30 seconds, then placed in the ultrasonic bath and sonicated (40 ± 2 kHz and 0.22 ± 0.04 W/cm 2 ) for 1 minute (4,20). The bottle was removed from the ultrasonic bath and vortexed again for 30 seconds.…”

Section: Ethical Approvalmentioning

confidence: 99%

“…The culture bottles were incubated in the automated BacT/Alert system. The VITEK 2 system (bioMérieux) was used for susceptibility testing (4,13,14).…”

Section: Ethical Approvalmentioning

confidence: 99%

“…However, the detection rate of the pathogen is low when a patient is receiving antibiotics (4). We focused on next-generation sequencing (NGS) to detect nonculturable microbes.…”

Section: Introductionmentioning

confidence: 99%

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Verification of a New Pathogen Detection Method by PostSonication Next-Generation Sequencing with Quantitative Real-Time PCR for the Diagnosis of Orthopedic Biofilm Infection

Ueda,

Inoue,

Okuda

et al. 2023

Preprint

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Next-generation sequencing (NGS), are recommended for the unconfirmed diagnosis of periprosthetic joint infection (PJI), but initial assessment using quantitative methods is required to determine whether microorganisms are present, before embarking on sequencing approaches. This study aimed to validate a new diagnostic approach for orthopedic biofilm infections that combines quantitative methods as quantitative real-time polymerase chain reaction (qPCR) with NGS. This study sequenced 16 clinical samples from one patient with PJI and 16 quantitative positive controls. The detection rate of multiple Staphylococcus species with NGS was approximately ten times higher than with culture. The read counts of Staphylococci detected in peri-implant samples were negatively correlated with the qPCR Cq value, while the three most commonly detected contaminant bacteria were positively correlated with the qPCR Cq values. The results were similar to those of the quantitative positive control study. In conclusion, In addition, pathogens and contaminants can also be discriminated by correlation with qPCR Cq values. These results of NGS with qPCR method can be used to determine the cause of clinically problematic infections in NGS that are difficult to confirm because false positive results in diagnosing infection are expected to cause misdiagnosis and misidentification of the causative organism, causing inappropriate therapy.

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Section: Ethical Approvalmentioning

confidence: 99%

“…The culture bottles were incubated in the automated BacT/Alert system. The VITEK 2 system (bioMérieux) was used for susceptibility testing (4,13,14).…”

Section: Ethical Approvalmentioning

confidence: 99%