ShoulderNeer Award 2017: A rapid method for detecting Propionibacterium acnes in surgical biopsy specimens from the shoulder
Section snippets
Primer design and enzyme digestion site identification
Genomic sequences of the 16S ribosomal RNA genes for a broad range of Propionibacterium spp, including P acnes, P avidum, P propionicus, P acidipropionici, P theonii, P granulosum, and P lymphophilum were obtained from the National Center for Biotechnology Information (NCBI, Bethesda, MD, USA). GenBank, the annotated collection of publicly available DNA sequences provided by NCBI, was used to obtain these Propionibacterium sequences in FASTA format, and they were entered into the European
PCR design, optimization, and validation
As described in the “Materials and methods,” comparative in silico analyses of the 16S rRNA gene sequences of 7 different Propionibacterium spp were performed to identify P acnes-specific targets for PCR amplification. One region of the 16S rRNA gene in P acnes that contained regions that differed from other closely related species and all other bacterial species is shown in Fig. 1. Some of these regions had been identified previously as targets for P acnes-specific PCR analyses in published
Discussion
There is an identified need for more rapid and accurate technologies to diagnose P acnes infections in patients after shoulder arthroplasty.11 The current standard for P acnes identification is a tissue swab for anaerobic culture,17 an approach that carries substantial risk of contamination from the adjacent skin and other sites where P acnes is a commensal organism. The length of time required for anaerobic culture and subsequent confirmatory analyses averages 6 or more days.17 In contrast,
Conclusion
The PCR-RFLP assay reported here is simple, robust, distinguishes between P acnes and other Propionibacterium spp, and is (theoretically) cost effective. In its current form, the assay provides proof of concept for using a PCR-RFLP approach to rapidly and consistently identify P acnes in tissues from shoulder biopsies within 24 hours. A potential additional advantage of this approach is that the minute sample size required for analysis may decrease the false-positive rate in cultures caused by
Disclaimer
Scott Holmes received financial support from the Schulich Research Opportunities Program in the Schulich School of Medicine and Dentistry at the University of Western Ontario. Kenneth J. Faber and David B. O'Gorman received support from the Department of Surgery at the University of Western Ontario through an Internal Research Fund award. George S. Athwal and David B. O'Gorman received a Canadian Orthopedics Research Legacy Award.
The authors, their immediate families, and any research
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2022, Journal of Molecular DiagnosticsCitation Excerpt :However, determination of cutoff values is challenging because there is no gold standard. Different methods are described in the literature,25,26,40,42–44 and PCR performances for cutibacteria vary.45–47 It was decided to determine cutoff values based on the PCR results of sterile samples (ie, culture-negative cerebrospinal fluid samples) because these samples are believed to be free of Cutibacterium background signal from colonization, and positive signals are expected only from contaminating Cutibacterium DNA in reagents.
Cutibacterium acnes phylogenetic type IC and II isolated from patients with non-acne diseases exhibit high-level biofilm formation
2021, International Journal of Medical MicrobiologyCitation Excerpt :Thus, C. acnes should be deemed as a pathogen for opportunistic infections. In the future, frequency of C. acnes detection may increase as causative pathogen of infectious diseases since a rapid detection method for C. acnes has been reported (Holmes et al., 2017). Proportion of high-BF strains isolated from non-acne patients was significantly higher than that of acne patients.
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The University of Western Ontario Research Ethics Board for Health Sciences Research Involving Human Subjects approved this study (No. 104888).