Multiplex Touchdown PCR for Rapid Typing of the Opportunistic Pathogen Propionibacterium acnes

Barnard, Emma; Nagy, István; Hunyadkürti, Judit; Patrick, Sheila; McDowell, Andrew

doi:10.1128/jcm.02460-14

Cited by 56 publications

(58 citation statements)

References 54 publications

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“…Using a previously described rapid multiplex PCR assay [36], we phylotyped all the isolates. We identified four different phylotypes: IA 1 (15 strains), IA 2 (2 strains), IB (9 strains) and II (2 strains) (Table 2, S3 Fig).…”

Section: Resultsmentioning

confidence: 99%

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TLR-2 Recognizes Propionibacterium acnes CAMP Factor 1 from Highly Inflammatory Strains

et al. 2016

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BackgroundPropionibacterium acnes (P. acnes) is an anaerobic, Gram-positive bacteria encountered in inflammatory acne lesions, particularly in the pilosebaceous follicle. P. acnes triggers a strong immune response involving keratinocytes, sebocytes and monocytes, the target cells during acne development. Lipoteicoic acid and peptidoglycan induce the inflammatory reaction, but no P. acnes surface protein interacting with Toll-like receptors has been identified. P. acnes surface proteins have been extracted by lithium stripping and shown to induce CXCL8 production by keratinocytes.Methodology and principal findingsFar-western blotting identified two surface proteins, of 24.5- and 27.5-kDa in size, specifically recognized by TLR2. These proteins were characterized, by LC-MS/MS, as CAMP factor 1 devoid of its signal peptide sequence, as shown by N-terminal sequencing. Purified CAMP factor 1 induces CXCL8 production by activating the CXCL8 gene promoter, triggering the synthesis of CXCL8 mRNA. Antibodies against TLR2 significantly decreased the CXCL8 response. For the 27 P. acnes strains used in this study, CAMP1-TLR2 binding intensity was modulated and appeared to be strong in type IB and II strains, which produced large amounts of CXCL8, whereas most of the type IA1 and IA2 strains presented little or no CAMP1-TLR2 binding and low levels of CXCL8 production. The nucleotide sequence of CAMP factor displays a major polymorphism, defining two distinct genetic groups corresponding to CAMP factor 1 with 14 amino-acid changes from strains phylotyped II with moderate and high levels of CAMP1-TLR2 binding activity, and CAMP factor 1 containing 0, 1 or 2 amino-acid changes from strains phylotyped IA1, IA2, or IB presenting no, weak or moderate CAMP1-TLR2 binding.ConclusionsOur findings indicate that CAMP factor 1 may contribute to P. acnes virulence, by amplifying the inflammation reaction through direct interaction with TLR2.

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Section: Resultsmentioning

confidence: 99%

“…Lanes 2 to 28: strains 12513, 14230, 16351, 17248, 22197, 22795, 25236, 27387, 27647, 38862, 41103, 47474, 53468, 75150, 78910, A9, A11, A24, A26, A30, A44, 6919, CHR, PIE, RON, TRI, and GUE, respectively. Letters a, b, c, and e correspond to amplicons of 677, 494, 145, and 351 bp, respectively, as previously described [36]. …”

Section: Supporting Informationmentioning

confidence: 99%

TLR-2 Recognizes Propionibacterium acnes CAMP Factor 1 from Highly Inflammatory Strains

et al. 2016

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“…Initial typing of multiple P. acnes isolates was performed using a rapid multiplex PCR assay as previously described30. This assay confirms P. acnes species identity and phylogeny of isolates (types IA 1 , IA 2 , IB and IC, II and III) based on primer sets targeting the 16S rRNA gene as well as a series of protein-encoding genes.…”

Section: Methodsmentioning

confidence: 95%

Strains of the Propionibacterium acnes type III lineage are associated with the skin condition progressive macular hypomelanosis

Barnard

Liu

Yankova

et al. 2016

Sci Rep

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Progressive macular hypomelanosis (PMH) is a common skin disorder that causes hypopigmentation in a variety of skin types. Although the underlying aetiology of this condition is unclear, there is circumstantial evidence that links the skin bacterium Propionibacterium acnes to the condition. We now describe the first detailed population genetic analysis of P. acnes isolates recovered from paired lesional and non-lesional skin of PMH patients. Our results demonstrate a strong statistical association between strains from the type III phylogenetic lineage and PMH lesions (P = 0.0019), but not those representing other phylogroups, including those associated with acne (type IA1). We also demonstrate, based on in silico 16S rDNA analysis, that PMH isolates previously recovered from patients in Europe are also consistent with the type III lineage. Using comparative genome analysis, we identified multiple genomic regions that are specific for, or absent from, type III strains compared to other phylogroups. In the former case, these include open reading frames with putative functions in metabolism, transport and transcriptional regulation, as well as predicted proteins of unknown function. Further study of these genomic elements, along with transcriptional and functional analyses, may help to explain why type III strains are associated with PMH.

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“…acnes isolates was purified using the QIAamp DNA mini kit (Qiagen). Phylotyping was conducted according to the MLST8 scheme and determined by a recently described multiplex PCR assay [15]. Refer to the cited publication for primer information and amplification conditions.…”

Section: Methodsmentioning

confidence: 99%

Propionibacterium acnes biofilm is present in intervertebral discs of patients undergoing microdiscectomy

et al. 2017

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BackgroundIn previous studies, Propionibacterium acnes was cultured from intervertebral disc tissue of ~25% of patients undergoing microdiscectomy, suggesting a possible link between chronic bacterial infection and disc degeneration. However, given the prominence of P. acnes as a skin commensal, such analyses often struggled to exclude the alternate possibility that these organisms represent perioperative microbiologic contamination. This investigation seeks to validate P. acnes prevalence in resected disc cultures, while providing microscopic evidence of P. acnes biofilm in the intervertebral discs.MethodsSpecimens from 368 patients undergoing microdiscectomy for disc herniation were divided into several fragments, one being homogenized, subjected to quantitative anaerobic culture, and assessed for bacterial growth, and a second fragment frozen for additional analyses. Colonies were identified by MALDI-TOF mass spectrometry and P. acnes phylotyping was conducted by multiplex PCR. For a sub-set of specimens, bacteria localization within the disc was assessed by microscopy using confocal laser scanning and FISH.ResultsBacteria were cultured from 162 discs (44%), including 119 cases (32.3%) with P. acnes. In 89 cases, P. acnes was cultured exclusively; in 30 cases, it was isolated in combination with other bacteria (primarily coagulase-negative Staphylococcus spp.) Among positive specimens, the median P. acnes bacterial burden was 350 CFU/g (12 - ~20,000 CFU/g). Thirty-eight P. acnes isolates were subjected to molecular sub-typing, identifying 4 of 6 defined phylogroups: IA1, IB, IC, and II. Eight culture-positive specimens were evaluated by fluorescence microscopy and revealed P. acnes in situ. Notably, these bacteria demonstrated a biofilm distribution within the disc matrix. P. acnes bacteria were more prevalent in males than females (39% vs. 23%, p = 0.0013).ConclusionsThis study confirms that P. acnes is prevalent in herniated disc tissue. Moreover, it provides the first visual evidence of P. acnes biofilms within such specimens, consistent with infection rather than microbiologic contamination.

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Multiplex Touchdown PCR for Rapid Typing of the Opportunistic Pathogen Propionibacterium acnes

Cited by 56 publications

References 54 publications

TLR-2 Recognizes Propionibacterium acnes CAMP Factor 1 from Highly Inflammatory Strains

TLR-2 Recognizes Propionibacterium acnes CAMP Factor 1 from Highly Inflammatory Strains

Strains of the Propionibacterium acnes type III lineage are associated with the skin condition progressive macular hypomelanosis

Propionibacterium acnes biofilm is present in intervertebral discs of patients undergoing microdiscectomy

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