Demonstration of Bacillus cereus in Orthopaedic-Implant-Related Infection with Use of a Multi-Primer Polymerase Chain Reaction-Mass Spectrometric Assay

Gallo, Phillip H.; Melton-Kreft, Rachael; Nistico, Laura; Sotereanos, Nicholas G.; Sewecke, Jeffrey J.; Stoodley, Paul; Ehrlich, Garth D.; Costerton, J. William; Kathju, Sandeep

doi:10.2106/jbjs.j.01181

Cited by 19 publications

(18 citation statements)

References 36 publications

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“…The partial concordance is attributable to the greater diversity found by the molecular methods vs. culture (Tables 2 and 3 and Fig. 1), which is consistent with similar comparative studies of other clinical conditions evaluating microbial detection methods [25, 44–48]. In the remaining three cases (4B, 7B and 8C) the disagreement between culture and molecular methods were caused by lack of culturability.…”

Section: Discussionsupporting

confidence: 84%

Comparing culture and molecular methods for the identification of microorganisms involved in necrotizing soft tissue infections

Rudkjøbing

Thomsen

et al. 2016

BMC Infect Dis

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BackgroundNecrotizing soft tissue infections (NSTIs) are a group of infections affecting all soft tissues. NSTI involves necrosis of the afflicted tissue and is potentially life threatening due to major and rapid destruction of tissue, which often leads to septic shock and organ failure. The gold standard for identification of pathogens is culture; however molecular methods for identification of microorganisms may provide a more rapid result and may be able to identify additional microorganisms that are not detected by culture.MethodsIn this study, tissue samples (n = 20) obtained after debridement of 10 patients with NSTI were analyzed by standard culture, fluorescence in situ hybridization (FISH) and multiple molecular methods. The molecular methods included analysis of microbial diversity by 1) direct 16S and D2LSU rRNA gene Microseq 2) construction of near full-length 16S rRNA gene clone libraries with subsequent Sanger sequencing for most samples, 3) the Ibis T5000 biosensor and 4) 454-based pyrosequencing. Furthermore, quantitative PCR (qPCR) was used to verify and determine the relative abundance of Streptococcus pyogenes in samples.ResultsFor 70 % of the surgical samples it was possible to identify microorganisms by culture. Some samples did not result in growth (presumably due to administration of antimicrobial therapy prior to sampling). The molecular methods identified microorganisms in 90 % of the samples, and frequently detected additional microorganisms when compared to culture. Although the molecular methods generally gave concordant results, our results indicate that Microseq may misidentify or overlook microorganisms that can be detected by other molecular methods.Half of the patients were found to be infected with S. pyogenes, but several atypical findings were also made including infection by a) Acinetobacter baumannii, b) Streptococcus pneumoniae, and c) fungi, mycoplasma and Fusobacterium necrophorum.ConclusionThe study emphasizes that many pathogens can be involved in NSTIs, and that no specific “NSTI causing” combination of species exists. This means that clinicians should be prepared to diagnose and treat any combination of microbial pathogens. Some of the tested molecular methods offer a faster turnaround time combined with a high specificity, which makes supplemental use of such methods attractive for identification of microorganisms, especially for fulminant life-threatening infections such as NSTI.

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Section: Discussionsupporting

confidence: 84%

Comparing culture and molecular methods for the identification of microorganisms involved in necrotizing soft tissue infections

Rudkjøbing

Thomsen

et al. 2016

BMC Infect Dis

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“…Dubouix et al [13] reported forty-one B. cereus infected cases, which were suspected of terrestrial environmental contamination before admission. Gallo et al [14] In conclusion, we report very late prosthetic joint infection at hip joint and consequent bacteremia by B. cereus, which occurred after 13 years post total hip replacement operation without recent history of trauma or intervention. This is rare case of very late prosthetic joint infection by B. cereus, necessitating caution to discriminate true infection from contamination of B.…”

Section: The Genus Bacillus Includes B Cereus B Mycoides B Pseudsupporting

confidence: 47%

Late Prosthetic Joint Infection and Bacteremia by Bacillus cereus Confirmed by 16S rRNA Sequencing and Hip Joint Tissue Pathology

Park

Kim

et al. 2016

Ann Clin Microbiol

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“…For the Ibis T5000 assay, an aliquot of each sample was loaded into each of the 16 wells of a 96-well bacterial artificial chromosome (BAC) detection PCR plate (PN 05N13-01; Abbott Molecular). Each of the 16 wells for each specimen contained a different broad-range primer set (14,24). An internal calibrant consisting of a synthetic nucleic acid template was included in each assay, controlling for false negatives (e.g., from PCR inhibitors) and enabling a semiquantitative analysis of the amount of template DNA present.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Bacterial Communities in Venous Insufficiency Wounds by Use of Conventional Culture and Molecular Diagnostic Methods

et al. 2011

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Microbial infections delay wound healing, but the effect of the composition of the wound microbiome on healing parameters is unknown. To better understand bacterial communities in chronic wounds, we analyzed debridement samples from lower-extremity venous insufficiency ulcers using the following: conventional anaerobic and aerobic bacterial cultures; the Ibis T5000 universal biosensor (Abbott Molecular); and 16S 454 FLX titanium series pyrosequencing (Roche). Wound debridement samples were obtained from 10 patients monitored clinically for at least 6 months, at which point 5 of the 10 sampled wounds had healed. Pyrosequencing data revealed significantly higher bacterial abundance and diversity in wounds that had not healed at 6 months. Additionally, Actinomycetales was increased in wounds that had not healed, and Pseudomonadaceae was increased in wounds that had healed by the 6-month follow-up. Baseline wound surface area, duration, or analysis by Ibis or conventional culture did not reveal significant differences between wounds that healed after 6 months and those that did not. Thus, pyrosequencing identified distinctive baseline characteristics of wounds that did not heal by the 6-month follow-up, furthering our understanding of potentially unique microbiome characteristics of chronic wounds.

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Demonstration of Bacillus cereus in Orthopaedic-Implant-Related Infection with Use of a Multi-Primer Polymerase Chain Reaction-Mass Spectrometric Assay

Cited by 19 publications

References 36 publications

Comparing culture and molecular methods for the identification of microorganisms involved in necrotizing soft tissue infections

Comparing culture and molecular methods for the identification of microorganisms involved in necrotizing soft tissue infections

Late Prosthetic Joint Infection and Bacteremia by Bacillus cereus Confirmed by 16S rRNA Sequencing and Hip Joint Tissue Pathology

Characterization of Bacterial Communities in Venous Insufficiency Wounds by Use of Conventional Culture and Molecular Diagnostic Methods

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