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Kim, Song, Lee, and Kim: Antimicrobial Susceptibility and Characterization of Propionibacterium acnes by Multilocus Sequence Typing and Repetitive-Sequence-Based PCR
Original Article

J Bacteriol Virol 2016;46(3):135-141.

Published online: 9 February 2016

DOI: https://doi.org/10.4167/jbv.2016.46.3.135

Antimicrobial Susceptibility and Characterization of Propionibacterium acnes by Multilocus Sequence Typing and Repetitive-Sequence-Based PCR

Shukho Kim1, Hyesoon Song1, Weon-Ju Lee2, Jungmin Kim1, ∗

1Department of Microbiology, Kyungpook National University School of Medicine, Daegu, Korea

2Department of Dermatology, Kyungpook National University School of Medicine, Daegu, Korea

∗Corresponding author: Jungmin Kim. Department of Microbiology, Kyungpook National University School of Medicine, 680 gukchaebosang-ro, Jung-gu, Daegu 41944, Korea. Phone: +82-53-420-4845, Fax: +82-53-427-5664, e-mail: minkim@knu.ac.kr

∗∗ The pathogens for this study were provided by the Kyungpook, Chonbuk, and Gyeongsang National University Hospital Culture Collections for Pathogens, members of the National Biobank of Korea are supported by the Ministry of Health and Welfare.

Received 2 August 201618 August 2016       Accepted 26 August 2016

Copyright © 2016 Journal of Bacteriology and Virology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Propionibacterium acnes, a gram-positive, anaerobic, and aerotolerant bacterium that is found frequently in the skin as part of the human microbiome causes inflammatory acne, shoulder infection, and the contamination of medical devices. The study goals were the antibiotic resistant and molecular epidemiological characterization of the P. acnes isolates in Korea. A total of 22 P. acnes isolates originated from diverse patients were obtained from three National Culture Collections for Pathogens in South Korea. The hemolytic properties and minimum inhibitory concentrations (MIC) of five antibiotics (tetracycline, doxycycline, clindamycin, erythromycin, and minocycline) were determined. Only one isolate showed high MIC values and resistance to all five antibiotics. Genotypic characterization was achieved by multilocus sequence typing (MLST) for eight loci (aroE, guaA, tly, camp2, atpD, gmk, lepA, and sodA) and repetitive-sequence-based PCR (rep-PCR) analysis using the DiversiLab kit. MLST revealed four phylogroups that were type IA1 (27.3%), type IA2 (18.2%), type IB (13.6%), and type II (40.9%). Rep-PCR results demonstrated three clusters that were cluster I (39.1%), cluster II (45.5%), and cluster III (13.6%). The isolates of cluster I were part of phylogroup type IA (both IA1 and IA2), and the isolates of cluster II belonged to phylogroup type II. All isolates of phylogroup type IB were hemolytic and belonged to cluster III. The results of rep-PCR clustering analysis showed a good correlation with those of MLST phylogroups, suggesting that rep-PCR could be an alternative method to track P. acnes subtype lineages.

Keywords: Propionibacterium acnes, Antimicrobial susceptibility, Multilocus sequence typing, rep-PCR

REFERENCES

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Figure 1.
Dendrogram and three similarity clusters generated from repetitive-sequence-based PCR results for 22 clinical isolates of P. acnes and one type strain, P. acnes ATCC 11828.
jbv-46-135f1.tif
Table 1.
Minimum inhibitory concentrations (µg/ml) of five antimicrobial agents using 22 clinical isolates and one standard strain of P. acnes
P. acnes isolate Tetracycline (RB1: 2.0) Doxycycline (RB: 1.0) Clindamycin (RB: 4.0) Erythromycin (RB: 0.5) Minocycline (RB: 1.0)
KB105 0.064 0.125 ≤ 0.016 ≤ 0.016 0.047
KB106 0.25 0.19 ≤ 0.016 ≤ 0.016 ≤ 0.016
KB109 0.25 0.38 0.023 ≤ 0.016 0.094
KB112 8 4 ≥ 256 ≥ 256 3
KB113 0.125 0.125 ≤ 0.016 0.023 0.047
KB114 0.25 0.25 ≤ 0.016 0.023 0.125
KB116 ≤ 0.016 0.19 ≤ 0.016 ≤ 0.016 ≤ 0.016
KB117 0.064 0.064 0.023 ≤ 0.016 0.023
KB118 0.064 0.032 ≤ 0.016 ≤ 0.016 0.125
KB119 0.5 0.25 ≤ 0.016 ≤ 0.016 0.047
JB391 0.125 0.047 ≤ 0.016 ≤ 0.016 0.023
JB893 0.25 0.5 0.023 ≤ 0.016 0.19
JB1475 0.125 0.094 ≤ 0.016 ≤ 0.016 0.047
JB1490 0.125 0.125 ≤ 0.016 ≤ 0.016 0.023
JB1912 0.25 0.25 0.023 ≤ 0.016 0.047
JB3672 0.047 0.032 ≤ 0.016 ≤ 0.016 ≤ 0.016
JB3496 0.064 0.064 ≤ 0.016 ≤ 0.016 0.023
KS2874 0.25 0.064 ≤ 0.016 ≤ 0.016 ≤ 0.016
KS2875 0.5 0.125 0.032 ≤ 0.016 0.094
KS2876 0.19 0.19 0.023 ≤ 0.016 0.047
KS2877 0.25 0.19 0.032 ≤ 0.016 0.032
KS2878 0.125 0.125 0.023 ≤ 0.016 ≤ 0.016
ATCC 11828 0.19 0.125 ≤ 0.016 ≤ 0.016 0.047

1 Resistance breakpoint according to the recommendation by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) for P. acnes (µg/ml).

Table 2.
Hemolysis, multilocus sequence typing (MLST), and repetitive-sequence-based PCR (rep-PCR) for 22 clinical isolates and 1 type strain of P. acnes
Isolate Hemolysis1 MLST Rep-PCR p
Allelic profile (aroE-guaA-tly-camp2-atpD- gmk-lepA-sodA) ST CC Phylogroup
KB105 - 1-5-8-2-1-1-1-4 2 CC2 IA2 Cluster I
KB106 - 1-5-8-7-1-1-1-5 22 Singleton IA2 Cluster I
KB109 + 1-3-1-1-1-1-1-1 1 CC1 IA1 Cluster I
KB112 + 1-3-22-2-1-1-1-1 115 CC3 IA1 Cluster I
KB113 + 1-4-8-6-1-9-1-4 53 CC5 IB Cluster III
KB114 + 1-5-8-7-1-1-1-5 22 Singleton IA2 Cluster I
KB116 - 17-4-10-12-4-2-4-6 69 CC72 II Cluster II
KB117 + 1-4-8-6-1-1-1-4 5 CC5 IB Cluster III
KB118 + 1-3-1-1-1-1-1-1 1 CC1 IA1 Cluster I
KB119 + 1-3-1-1-1-1-1-1 1 CC1 IA1 Cluster I
JB391 + 17-4-10-12-4-2-4-6 69 CC72 II Cluster II
JB893 + 1-3-1-1-1-1-1-1 1 CC1 IA1 Cluster I
JB1475 - 17-4-10-12-4-2-4-6 69 CC72 II Cluster II
JB1490 - 17-4-10-10-4-2-2-3 6 CC6 II Cluster II
JB1912 - 17-4-10-10-4-2-2-3 6 CC6 II Cluster II
JB3496 + 1-3-22-2-1-1-1-1 115 CC3 IA1 Cluster I
JB3672 - 17-4-10-12-4-2-4-6 69 CC72 II Cluster II
KS2874 - 1-5-8-2-1-1-1-4 2 CC2 IA2 Cluster I
KS2875 - 17-4-10-10-4-2-2-3 6 CC6 II Cluster II
KS2876 - 17-4-10-12-4-2-4-6 69 CC72 II Cluster II
KS2877 - 17-4-10-10-9-2-2-3 25 CC6 II Cluster II
KS2878 + 1-4-8-6-1-9-1-4 53 CC5 IB Cluster III
ATCC 11828 - 17-4-10-12-4-2-4-6 69 CC72 II Cluster I

1 +: hemolysis positive, -: hemolysis negative.

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