Volume 26, Number 10—October 2020
Dispatch
Limitations of Ribotyping as Genotyping Method for Corynebacterium ulcerans
Figure 1

Figure 1. Alteration of ribotyping patterns by genomic DNA modification of Corynebacterium ulcerans strains 0102, 0211, and FH2016–1, Japan, 2001–2016. Ribotyping was performed as described previously (4,11). HindIII-digested, digoxigenin-labeled λ phage DNA segments were used as length markers. A) Conventional ribotyping patterns of strains 0102, 0211, and FH2016-1. 1, λHindIII; 2, 0102; 3, 0211; 4, FH2016-1; 5, Pattern predicted by in silico typing. B) Ribotyping patterns of genomic DNA and whole-genome amplified DNA as substrates. 1, λHindIII; 2, 0102 WGA; 3, 0102 native; 4, 0211 WGA; 5, 0211 native; 6, FH2016-1 WGA; 7, FH2016-1 native. The label “WGA” indicates whole-genome amplified DNA as a substrate; “native” indicates genomic DNA. WGA (unmodified) DNA of the 3 strains show identical patterns. The pattern matches that of native 0211 (unmodified genomic DNA). In contrast, native FH2016-1 and 0102 are modified and show different patterns from their WGA counterparts.
References
- World Health Organization. Diphtheria. In: Vaccine preventable diseases surveillance standards. Geneva: The Organization; 2018.
- De Zoysa A, Hawkey P, Charlett A, Efstratiou A. Comparison of four molecular typing methods for characterization of Corynebacterium diphtheriae and determination of transcontinental spread of C. diphtheriae based on BstEII rRNA gene profiles. J Clin Microbiol. 2008;46:3626–35. DOIPubMedGoogle Scholar
- Grimont PAD, Grimont F, Efstratiou A, De Zoysa A, Mazurova I, Ruckly C, et al.; European Laboratory Working Group on Diphtheria. International nomenclature for Corynebacterium diphtheriae ribotypes. Res Microbiol. 2004;155:162–6. DOIPubMedGoogle Scholar
- De Zoysa A, Hawkey PM, Engler K, George R, Mann G, Reilly W, et al. Characterization of toxigenic Corynebacterium ulcerans strains isolated from humans and domestic cats in the United Kingdom. J Clin Microbiol. 2005;43:4377–81. DOIPubMedGoogle Scholar
- Komiya T, Seto Y, De Zoysa A, Iwaki M, Hatanaka A, Tsunoda A, et al. Two Japanese Corynebacterium ulcerans isolates from the same hospital: ribotype, toxigenicity and serum antitoxin titre. J Med Microbiol. 2010;59:1497–504. DOIPubMedGoogle Scholar
- Otsuji K, Fukuda K, Endo T, Shimizu S, Harayama N, Ogawa M, et al. The first fatal case of Corynebacterium ulcerans infection in Japan. JMM Case Rep. 2017;4:
e005106 . DOIPubMedGoogle Scholar - Yasuda I, Matsuyama H, Ishifuji T, Yamashita Y, Takaki M, Morimoto K, et al. Severe pneumonia caused by toxigenic Corynebacterium ulcerans infection, Japan. Emerg Infect Dis. 2018;24:588–91. DOIPubMedGoogle Scholar
- Katsukawa C, Komiya T, Umeda K, Goto M, Yanai T, Takahashi M, et al. Toxigenic Corynebacterium ulcerans isolated from a hunting dog and its diphtheria toxin antibody titer. Microbiol Immunol. 2016;60:177–86. DOIPubMedGoogle Scholar
- Katsukawa C, Umeda K, Inamori I, Kosono Y, Tanigawa T, Komiya T, et al. Toxigenic Corynebacterium ulcerans isolated from a wild bird (ural owl) and its feed (shrew-moles): comparison of molecular types with human isolates. BMC Res Notes. 2016;9:181. DOIPubMedGoogle Scholar
- König C, Meinel DM, Margos G, Konrad R, Sing A. Multilocus sequence typing of Corynebacterium ulcerans provides evidence for zoonotic transmission and for increased prevalence of certain sequence types among toxigenic strains. J Clin Microbiol. 2014;52:4318–24. DOIPubMedGoogle Scholar
- Hatanaka A, Tsunoda A, Okamoto M, Ooe K, Nakamura A, Miyakoshi M, et al. Corynebacterium ulcerans Diphtheria in Japan. Emerg Infect Dis. 2003;9:752–3. DOIPubMedGoogle Scholar
- Sekizuka T, Yamamoto A, Komiya T, Kenri T, Takeuchi F, Shibayama K, et al. Corynebacterium ulcerans 0102 carries the gene encoding diphtheria toxin on a prophage different from the C. diphtheriae NCTC 13129 prophage. BMC Microbiol. 2012;12:72. DOIPubMedGoogle Scholar
- Regnault B, Grimont F, Grimont PAD. Universal ribotyping method using a chemically labelled oligonucleotide probe mixture. Res Microbiol. 1997;148:649–59. DOIPubMedGoogle Scholar
- Roberts RJ, Vincze T, Posfai J, Macelis D. REBASE—a database for DNA restriction and modification: enzymes, genes and genomes. Nucleic Acids Res. 2015;43(D1):D298–9. DOIPubMedGoogle Scholar
- Nelson JR. Random-primed, Phi29 DNA polymerase-based whole genome amplification. Curr Protoc Mol Biol. 2014;105:15.13.1.
1Current affiliation: Osaka Prefecture University, Osaka, Japan.