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Volume 26, Number 10—October 2020
Dispatch

Limitations of Ribotyping as Genotyping Method for Corynebacterium ulcerans

Tsuyoshi Sekizuka, Chihiro Katsukawa1, Makoto Kuroda, Keigo Shibayama, Ken Otsuji, Mitsumasa Saito, Akihiko Yamamoto, and Masaaki IwakiComments to Author 
Author affiliations: National Institute of Infectious Diseases, Tokyo, Japan (T. Sekizuka, M. Kuroda, K. Shibyama, A. Yamamoto, M. Iwaki); Osaka Prefectural Institute of Public Health, Osaka, Japan (C. Katsukawa); University of Occupational and Environmental Health, Kitakyushu, Japan (K. Otsuji, M. Saito)

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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.

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.

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References
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1Current affiliation: Osaka Prefecture University, Osaka, Japan.

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