Ann Lab Med.  2019 Nov;39(6):530-536. 10.3343/alm.2019.39.6.530.

Application of the Whole Genome-Based Bacterial Identification System, TrueBac ID, Using Clinical Isolates That Were Not Identified With Three Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) Systems

Affiliations
  • 1ChunLab, Inc., Seoul, Korea.
  • 2School of Biological Sciences & Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Korea.
  • 3Seoul Clinical Laboratories, Yongin, Korea.
  • 4Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea. deyong@yuhs.ac
  • 5Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea.
  • 6Department of Laboratory Medicine, Gyeongsang National University Hospital, and Gyeongsang National University College of Medicine, Jinju, Korea.
  • 7Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.

Abstract

BACKGROUND
Next-generation sequencing is increasingly used for taxonomic identification of pathogenic bacterial isolates. We evaluated the performance of a newly introduced whole genome-based bacterial identification system, TrueBac ID (ChunLab Inc., Seoul, Korea), using clinical isolates that were not identified by three matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems and 16S rRNA gene sequencing.
METHODS
Thirty-six bacterial isolates were selected from a university-affiliated hospital and a commercial clinical laboratory. Species was identified by three MALDI-TOF MS systems: Bruker Biotyper MS (Bruker Daltonics, Billerica, MA, USA), VITEK MS (bioMérieux, Marcy l'Étoile, France), and ASTA MicroIDSys (ASTA Inc., Suwon, Korea). Whole genome sequencing was conducted using the Illumina MiSeq system (Illumina, San Diego, CA, USA), and genome-based identification was performed using the TrueBac ID cloud system (www.truebacid.com).
RESULTS
TrueBac ID assigned 94% (34/36) of the isolates to known (N=25) or novel (N=4) species, genomospecies (N=3), or species group (N=2). The remaining two were identified at the genus level.
CONCLUSIONS
TrueBac ID successfully identified the majority of isolates that MALDI-TOF MS failed to identify. Genome-based identification can be a useful tool in clinical laboratories, with its superior accuracy and database-driven operations.

Keyword

Next generation sequencing; Genome-based identification; TrueBac ID; Performance

MeSH Terms

Genes, rRNA
Genome
Gyeonggi-do
Mass Spectrometry*
Seoul

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Reference

1. Vouga M, Greub G. Emerging bacterial pathogens: the past and beyond. Clin Microbiol Infect. 2016; 22:12–21. PMID: 26493844.
2. Croxatto A, Prod'hom G, Greub G. Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology. FEMS Microbiol Rev. 2012; 36:380–407. PMID: 22092265.
3. Van den Abeele AM, Vogelaers D, Vandamme P, Vanlaere E, Houf K. Filling the gaps in clinical proteomics: a do-it-yourself guide for the identification of the emerging pathogen Arcobacter by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Microbiol Methods. 2018; 152:92–97. PMID: 30017851.
4. Chun J, Rainey FA. Integrating genomics into the taxonomy and systematics of the Bacteria and Archaea. Int J Syst Evol Microbiol. 2014; 64:316–324. PMID: 24505069.
5. Yoon SH, Ha SM, Kwon S, Lim J, Kim Y, Seo H, et al. Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol. 2017; 67:1613–1617. PMID: 28005526.
6. Kim M, Oh HS, Park SC, Chun J. Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. Int J Syst Evol Microbiol. 2014; 64:346–351. PMID: 24505072.
7. Fox GE, Wisotzkey JD, Jurtshuk PJ Jr. How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. Int J Syst Bacteriol. 1992; 42:166–170. PMID: 1371061.
8. Chun J, Oren A, Ventosa A, Christensen H, Arahal DR, da Costa MS, et al. Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol. 2018; 68:461–466. PMID: 29292687.
9. Salipante SJ, SenGupta DJ, Cummings LA, Land TA, Hoogestraat DR, Cookson BT. Application of whole-genome sequencing for bacterial strain typing in molecular epidemiology. J Clin Microbiol. 2015; 53:1072–1079. PMID: 25631811.
10. Roach DJ, Burton JN, Lee C, Stackhouse B, Butler-Wu SM, Cookson BT, et al. A year of infection in the Intensive Care Unit: prospective whole genome sequencing of bacterial clinical isolates reveals cryptic transmissions and novel microbiota. PLoS Genet. 2015; 11:e1005413. PMID: 26230489.
11. Mellmann A, Bletz S, Böking T, Kipp F, Becker K, Schultes A, et al. Real-Time Genome sequencing of resistant bacteria provides precision infection control in an institutional setting. J Clin Microbiol. 2016; 54:2874–2881. PMID: 27558178.
12. Kyrpides NC, Hugenholtz P, Eisen JA, Woyke T, Göker M, Parker CT, et al. Genomic encyclopedia of bacteria and archaea: sequencing a myriad of type strains. PLoS Biol. 2014; 12:e1001920. PMID: 25093819.
13. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014; 30:2114–2120. PMID: 24695404.
14. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M, Kulikov AS, et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol. 2012; 19:455–477. PMID: 22506599.
15. Richter M, Rosselló-Móra R. Shifting the genomic gold standard for the prokaryotic species definition. Proc Natl Acad Sci U S A. 2009; 106:19126–19131. PMID: 19855009.
16. Murray RG, Stackebrandt E. Taxonomic note: implementation of the provisional status Candidatus for incompletely described procaryotes. Int J Syst Bacteriol. 1995; 45:186–187. PMID: 7857801.
17. Fischer S, Mayer-Scholl A, Imholt C, Spierling NG, Heuser E, Schmidt S, et al. Leptospira genomospecies and sequence type prevalence in small mammal populations in Germany. Vector Borne Zoonotic Dis. 2018; 18:188–199. PMID: 29470107.
18. Patil PP, Kumar S, Midha S, Gautam V, Patil PB. Taxonogenomics reveal multiple novel genomospecies associated with clinical isolates of Stenotrophomonas maltophilia. Microb Genom. 2018; 4:e000207.
19. Salipante SJ, Kalapila A, Pottinger PS, Hoogestraat DR, Cummings L, Duchin JS, et al. Characterization of a multidrug-resistant, novel Bacteroides genomospecies. Emerg Infect Dis. 2015; 21:95–98. PMID: 25529016.
20. Na SI, Kim YO, Yoon SH, Ha SM, Baek I, Chun J. UBCG: Up-to-date bacterial core gene set and pipeline for phylogenomic tree reconstruction. J Microbiol. 2018; 56:280–285. PMID: 29492869.
21. Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, et al. Introducing Ez-Taxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol. 2012; 62:716–721. PMID: 22140171.
22. Ondov BD, Treangen TJ, Melsted P, Mallonee AB, Bergman NH, Koren S, et al. Mash: fast genome and metagenome distance estimation using MinHash. Genome Biol. 2016; 17:132. PMID: 27323842.
23. CLSI. Interpretive criteria for identification of bacteria and fungi by DNA target sequencing. Approved guideline MM18-A. Wayne, PA: Clinical and Laboratory Standards Institute;2008.
24. Jang KS, Kim YH. Rapid and robust MALDI-TOF MS techniques for microbial identification: a brief overview of their diverse applications. J Microbiol. 2018; 56:209–216. PMID: 29492868.
25. Ndongo S, Andrieu C, Fournier PE, Lagier JC, Raoult D. ‘Actinomyces provencensis’ sp. nov., ‘Corynebacterium bouchesdurhonense’ sp. nov., ‘Corynebacterium provencense’ sp. nov. and ‘Xanthomonas massiliensis’ sp. nov., 4 new species isolated from fresh stools of obese French patients. New Microbes New Infect. 2017; 18:24–27. PMID: 28507764.
26. Kittl S, Brodard I, Rychener L, Jores J, Roosje P, Gobeli Brawand S. Otitis in a cat associated with Corynebacterium provencense. BMC Vet Res. 2018; 14:200. PMID: 29940943.
27. Fonkou MDM, Bilen M, Cadoret F, Fournier PE, Dubourg G, Raoult D. ‘Enterococcus timonensis’ sp. nov., ‘Actinomyces marseillensis’ sp. nov., ‘Leptotrichia massiliensis’ sp. nov., ‘Actinomyces pacaensis’ sp. nov., ‘Actinomyces oralis’ sp. nov., ‘Actinomyces culturomici’ sp. nov. and ‘Gemella massiliensis’ sp. nov., new bacterial species isolated from the human respiratory microbiome. New Microbes New Infect. 2017; 22:37–43. PMID: 29556407.
28. Martiny D, Visscher A, Catry B, Chatellier S, Vandenberg O. Optimization of Campylobacter growth conditions for further identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). J Microbiol Methods. 2013; 94:221–223. PMID: 23811211.
29. Mellmann A, Cloud J, Maier T, Keckevoet U, Ramminger I, Iwen P, et al. Evaluation of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry in comparison to 16S rRNA gene sequencing for species identification of nonfermenting bacteria. J Clin Microbiol. 2008; 46:1946–1954. PMID: 18400920.
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