Journal List > J Bacteriol Virol > v.46(4) > 1034223

J Bacteriol Virol. 2016 Dec;46(4):201-212. Korean.
Published online December 31, 2016.  https://doi.org/10.4167/jbv.2016.46.4.201
Copyright © 2016 The Korean Society for Microbiology and The Korean Society of Virology
Resource Development and Investigation of Novel Species from Unidentified Pathogens in NCCP using MALDI-TOF MS and 16S rRNA Gene Analysis
Won Seon Yu, Kyeong Min Lee and Kyu Jam Hwang
Pathogen Resource TF, Center for Infectious Diseases, Korea National Institute of Health, Korea Center for Disease Control and Prevention, Cheongju, Korea.

Corresponding author: Kyu Jam Hwang, Ph.D. Pathogen Resource TF, Center for Infectious Diseases, Korea National Institute of Health, Korea Center for Disease Control and Prevention, Cheongju, 28159, Korea. Phone: +82-43-719-6870, Fax: +82-43-719-6680, Email: kyuhwang@nih.go.kr
Received October 12, 2016; Revised November 21, 2016; Accepted November 25, 2016.

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/).


Abstract

Species identification is an important item to characterize unidentified bacterial pathogens in developing and managing bacterial resources. In this study, unidentified pathogens based on the results of an automated identification system were identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALD-TOF MS) and 16S rRNA gene analysis for development of national resources in the National Culture Collection for Pathogens (NCCP) in Korea. A total of 437 unidentified strains from branch banks of the NCCP were collected, and 16S rRNA and dnaJ gene sequencing, as well as MALDI-TOF MS analysis were performed to identify bacterial species. The mass spectra extracted were analyzed. Twelve strains exhibiting less than 98.65% similarity in 16S rRNA gene were selected as the primary candidates for novel species, and 21 strains exhibiting 98.65~99.0% similarity in 16S rRNA gene were selected as possible candidates for novel species. Among them, strain 32, belonging to Dermabacter sp., was finally selected as a possible strain representing a novel species and 14 unidentified bacterial strains using automated phenotypic identification system were newly registered at NCCP. The present study showed that unidentified pathogens using the automated phenotypic identification system were efficiently identified using the combination of MALDI-TOF MS and 16S rRNA gene analysis, and developed to the national resources in NCCP.

Keywords: Identification; MALDI-TOF MS; 16S rRNA; Pathogen; Resource

Figures


Figure 1
Phylogenetic tree based on the 16S rRNA gene sequences of strain 32.
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Figure 2
Phylogenetic tree based on the 16S rRNA gene sequences of strain 161.
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Figure 3
Phylogenetic tree based on the dnaJ gene sequence of strain 161.
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Figure 4
The modified phylogenetic tree based on the 16S rRNA gene of strain 161.
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Figure 5
Schematic representation of single major spectrometric profile from strain 32 (a) and Dermabacter hominis ATCC49369 (b).
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Figure 6
Dendrogram based on major spectrum profile of Dermabacteraceae.
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Tables


Table 1
Clinical information of newly-registered bacterial resources
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Table 2
Identification results of strain 32 and Dermabacter hominis ATCC49369 using MALDI-TOF MS and 16S rRNA gen
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Table 3
Biochemical characteristics of strain 32
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Notes

Bacterial strains used in this study were obtained from designated banks for culture collection. This research was supported and funded by Korea National Institute of Health (2013-NG45006-00).

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