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

Barnard, Emma, Nagy, Istvan, Hunyadkurti, Judit, Patrick, Sheila and McDowell, Andrew (2015) Multiplex Touchdown PCR for Rapid Typing of the Opportunistic Pathogen Propionibacterium acnes. Journal of Clinical Microbiology, 53 (4). pp. 1149-1155. [Journal article]

[img] Text - Accepted Version

DOI: 10.1128/JCM.02460-14


The opportunistic human pathogen Propionibacterium acnes is composed of a number of distinct phylogroups, designated types IA1, IA2, IB, IC, II, and III, which vary in their production of putative virulence factors, their inflammatory potential, and their biochemical, aggregative, and morphological characteristics. Although multilocus sequence typing (MLST) currently represents the gold standard for unambiguous phylogroup classification and individual strain identification, it is a labor-intensive and time-consuming technique. As a consequence, we developed a multiplex touchdown PCR assay that in a single reaction can confirm the species identity and phylogeny of an isolate based on its pattern of reaction with six primer sets that target the 16S rRNA gene (all isolates), ATPase (types IA1, IA2, and IC), sodA (types IA2 and IB), atpD (type II), and recA (type III) housekeeping genes, as well as a Fic family toxin gene (type IC). When applied to 312 P. acnes isolates previously characterized by MLST and representing types IA1 (n=145), IA2 (n=20), IB (n=65), IC (n=7), II (n=45), and III (n=30), the multiplex displayed 100% sensitivity and 100% specificity for detecting isolates within each targeted phylogroup. No cross-reactivity with isolates from other bacterial species was observed. This multiplex assay will provide researchers with a rapid, high-throughput, and technically undemanding typing method for epidemiological and phylogenetic investigations. It will facilitate studies investigating the association of lineages with various infections and clinical conditions, and it will serve as a prescreening tool to maximise the number of genetically diverse isolates selected for downstream higher-resolution sequence-based analyses.

Item Type:Journal article
Keywords:Propionibacterium acnes, multiplex PCR, phylogroups
Faculties and Schools:Faculty of Life and Health Sciences > School of Biomedical Sciences
Faculty of Life and Health Sciences
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Stratified Medicine
ID Code:34628
Deposited By: Dr Andrew McDowell
Deposited On:23 May 2016 15:02
Last Modified:17 Oct 2017 16:23

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