Analysis of clinical isolates of<i>Propionibacterium acnes</i>by optimised RAPD

Perry, Alexandra L.; Worthington, Tony; Hilton, Anthony C.; Lambert, Peter A.; Stirling, Allan; Elliott, Tom

doi:10.1016/s0378-1097(03)00720-1

Cited by 34 publications

(30 citation statements)

References 16 publications

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“…Such genes are under selective pressures different from those for universal housekeeping genes and therefore may be less well conserved, giving rise to better phylogenetic resolution of closely related organisms. Recently, analysis of 46 P. acnes isolates by random amplification of polymorphic DNA (RAPD) revealed two distinct genomic profiles (28). Although the type status of these various isolates was not determined, RAPD profiles from the type I and II reference strains, NCTC 737 and NCTC 10390, respectively, did match the two RAPD lineages of these P. acnes isolates, providing further evidence for the distinct genomic natures of types I and II.…”

Section: Discussionmentioning

confidence: 99%

Propionibacterium acnes Types I and II Represent Phylogenetically Distinct Groups

et al. 2005

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Although two phenotypes of the opportunistic pathogen Propionibacterium acnes (types I and II) have been described, epidemiological investigations of their roles in different infections have not been widely reported. Using immunofluorescence microscopy with monoclonal antibodies (MAbs) QUBPa1 and QUBPa2, specific for types I and II, respectively, we investigated the prevalences of the two types among 132 P. acnes isolates. Analysis of isolates from failed prosthetic hip implants (n ‫؍‬ 40) revealed approximately equal numbers of type I and II organisms. Isolates from failed prosthetic hip-associated bone (n ‫؍‬ 6) and tissue (n ‫؍‬ 38) samples, as well as isolates from acne (n ‫؍‬ 22), dental infections (n ‫؍‬ 8), and skin removed during surgical incision (n ‫؍‬ 18) were predominately of type I. A total of 11 (8%) isolates showed atypical MAb labeling and could not be conclusively identified. Phylogenetic analysis of P. acnes by nucleotide sequencing revealed the 16S rRNA gene to be highly conserved between types I and II. In contrast, sequence analysis of recA and a putative hemolysin gene (tly) revealed significantly greater type-specific polymorphisms that corresponded to phylogenetically distinct cluster groups. All 11 isolates with atypical MAb labeling were identified as type I by sequencing. Within the recA and tly phylogenetic trees, nine of these isolates formed a cluster distinct from other type I organisms, suggesting a further phylogenetic subdivision within type I. Our study therefore demonstrates that the phenotypic differences between P. acnes types I and II reflect deeper differences in their phylogeny. Furthermore, nucleotide sequencing provides an accurate method for identifying the type status of P. acnes isolates.Propionibacterium acnes is an opportunistic pathogen implicated in late-stage prosthetic joint infections, acne vulgaris, endocarditis, endophthalmitis, osteomyelitis, and shunt-associated central nervous system infections (2,5,7,33). Currently, routine diagnostic practices may underestimate the clinical importance of this anaerobic organism due to inefficient detection and isolation procedures, along with the traditionally held view that, due to its low virulence, its presence in clinical samples reflects contamination. While the opportunistic pathogenic potential of coagulase-negative staphylococci (CoNS), such as Staphylococcus epidermidis, is well recognized, the importance of P. acnes may still be overlooked (13), despite the fact that it produces more kinds of putative virulence determinants than CoNS (5, 38). This fact is illustrated by recent studies in which P. acnes was recovered as frequently as CoNS from the prosthetic hips of patients undergoing revision arthroplasty (33, 34).As a member of the resident human microbiota, P. acnes is found predominantly in the sebaceous gland-rich areas of the skin in adults (5, 25). It has, however, also been isolated from the conjunctiva, the mouth, and the large intestine (7). It accounts for approximately half of the total skin microb...

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Section: Discussionmentioning

confidence: 99%

Propionibacterium acnes Types I and II Represent Phylogenetically Distinct Groups

et al. 2005

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“…Index of association values (I A ) were determined following the method described by Haubold & Hudson (2000) with LIAN v. 3.5 software. Clonal groups were identified using the eBURST v. 3 clustering algorithm which does not impose a tree-like pattern of descent and dRAPD analysis as described by Perry et al (2003). §eBURST analysis following the method described by Feil et al (2004).…”

Section: Methodsmentioning

confidence: 99%

A novel multilocus sequence typing scheme for the opportunistic pathogen Propionibacterium acnes and characterization of type I cell surface-associated antigens

et al. 2011

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We have developed a novel multilocus sequence typing (MLST) scheme and database (http://pubmlst. org/pacnes/) for Propionibacterium acnes based on the analysis of seven core housekeeping genes. The scheme, which was validated against previously described antibody, single locus and random amplification of polymorphic DNA typing methods, displayed excellent resolution and differentiated 123 isolates into 37 sequence types (STs). An overall clonal population structure was detected with six eBURST groups representing the major clades I, II and III, along with two singletons. Two highly successful and global clonal lineages, ST6 (type IA) and ST10 (type IB 1 ), representing 64 % of this current MLST isolate collection were identified. The ST6 clone and closely related single locus variants, which comprise a large clonal complex CC6, dominated isolates from patients with acne, and were also significantly associated with ophthalmic infections. Our data therefore support an association between acne and P. acnes strains from the type IA cluster and highlight the role of a widely disseminated clonal genotype in this condition. Characterization of type I cell surface-associated antigens that are not detected in ST10 or strains of type II and III identified two dermatan-sulphate-binding proteins with putative phase/antigenic variation signatures. We propose that the expression of these proteins by type IA organisms contributes to their role in the pathophysiology of acne and helps explain the recurrent nature of the disease. The MLST scheme and database described in this study should provide a valuable platform for future epidemiological and evolutionary studies of P. acnes.

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“…Magnesium chloride, an important cofactor for Taq DNA Polymerase and for primer-template binding, is another important factor that influences reproducibility (Perry et al 2003). It was reasoned that a standard, relatively low, fixed concentration of 1.5 mM would minimize the likelihood of Fig.…”

Section: Primer Design and Pcr Protocolmentioning

confidence: 99%

Start Codon Targeted (SCoT) Polymorphism: A Simple, Novel DNA Marker Technique for Generating Gene-Targeted Markers in Plants

2008

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Random amplified polymorphic DNA (RAPD) markers have been used for numerous applications in plant molecular genetics research despite having disadvantages of poor reproducibility and not generally being associated with gene regions. A novel method for generating plant DNA markers was developed based on the short conserved region flanking the ATG start codon in plant genes. This method uses single 18-mer primers in single primer polymerase chain reaction (PCR) and an annealing temperature of 50°C. PCR amplicons are resolved using standard agarose gel electrophoresis. This method was validated in rice using a genetically diverse set of genotypes and a backcross population. Reproducibility was evaluated by using duplicate samples and conducting PCR on different days. Start codon targeted (SCoT) markers were generally reproducible but exceptions indicated that primer length and annealing temperature are not the sole factors determining reproducibility. SCoT marker PCR amplification profiles indicated dominant marker like RAPD markers. We propose that this method could be used in conjunction with these markers for applications such as genetic analysis, bulked segregant analysis, and quantitative trait loci mapping, especially in laboratories with a preference for agarose gel electrophoresis.

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Analysis of clinical isolates ofPropionibacterium acnesby optimised RAPD

Cited by 34 publications

References 16 publications

Propionibacterium acnes Types I and II Represent Phylogenetically Distinct Groups

Propionibacterium acnes Types I and II Represent Phylogenetically Distinct Groups

A novel multilocus sequence typing scheme for the opportunistic pathogen Propionibacterium acnes and characterization of type I cell surface-associated antigens

Start Codon Targeted (SCoT) Polymorphism: A Simple, Novel DNA Marker Technique for Generating Gene-Targeted Markers in Plants

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