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Abstract

Core genome multilocus sequence typing (cgMLST) has gained in popularity for bacterial typing since whole-genome sequencing (WGS) has become affordable. We introduce here pyMLST, a new complete, stand-alone, free and open source pipeline for cgMLST analysis. pyMLST can create or import a core genome database. For each gene, the first allele is aligned against the bacterial genome of interest using BLAT. Incomplete genes are aligned using MAFT. All data are stored in a SQLite database. pyMLST accepts assembly genomes or raw data (with the option pyMLST-KMA) as input. To evaluate our new tool, we selected three genome collections of major bacterial pathogens (, and ) and compared them with pyMLST, pyMLST-KMA, ChewBBACA, SeqSphere and the variant calling approach. We compared the sensitivity, precision and false-positive rate for each method with those of the variant calling approach. Minimal spanning trees were generated with each type of software to evaluate their interest in the context of a bacterial outbreak. We found that pyMLST-KMA is a convenient screening method to avoid assembling large bacterial collections. Our data showed that pyMLST (free, open source, available in Galaxy and pipeline ready) performed similarly to the commercial SeqSphere and performed better than ChewBBACA and pyMLST-KMA.

Keyword(s): bacteria , cgMLST , pyMLST , software and typing
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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/content/journal/mgen/10.1099/mgen.0.001126
2023-11-15
2024-09-14
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References

  1. Neoh H-M, Tan X-E, Sapri HF, Tan TL. Pulsed-field gel electrophoresis (PFGE): a review of the “gold standard” for bacteria typing and current alternatives. Infect Genet Evol 2019; 74:103935 [View Article] [PubMed]
    [Google Scholar]
  2. Besser J, Carleton HA, Gerner-Smidt P, Lindsey RL, Trees E. Next-generation sequencing technologies and their application to the study and control of bacterial infections. Clin Microbiol Infect 2018; 24:335–341 [View Article] [PubMed]
    [Google Scholar]
  3. Martak D, Meunier A, Sauget M, Cholley P, Thouverez M et al. Comparison of pulsed-field gel electrophoresis and whole-genome-sequencing-based typing confirms the accuracy of pulsed-field gel electrophoresis for the investigation of local Pseudomonas aeruginosa outbreaks. J Hosp Infect 2020; 105:643–647 [View Article] [PubMed]
    [Google Scholar]
  4. Pearce ME, Alikhan N-F, Dallman TJ, Zhou Z, Grant K et al. Comparative analysis of core genome MLST and SNP typing within a European Salmonella serovar Enteritidis outbreak. Int J Food Microbiol 2018; 274:1–11 [View Article] [PubMed]
    [Google Scholar]
  5. Silva M, Machado MP, Silva DN, Rossi M, Moran-Gilad J et al. chewBBACA: a complete suite for gene-by-gene schema creation and strain identification. Microb Genom 2018; 4:e000166 [View Article] [PubMed]
    [Google Scholar]
  6. Feijao P, Yao H-T, Fornika D, Gardy J, Hsiao W et al. MentaLiST - a fast MLST caller for large MLST schemes. Microb Genom 2018; 4:e000146 [View Article] [PubMed]
    [Google Scholar]
  7. Kent WJ. BLAT--the BLAST-like alignment tool. Genome Res 2002; 12:656–664 [View Article] [PubMed]
    [Google Scholar]
  8. Clausen P, Aarestrup FM, Lund O. Rapid and precise alignment of raw reads against redundant databases with KMA. BMC Bioinformatics 2018; 19:307 [View Article] [PubMed]
    [Google Scholar]
  9. Katoh K, Rozewicki J, Yamada KD. MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Brief Bioinform 2019; 20:1160–1166 [View Article] [PubMed]
    [Google Scholar]
  10. Blankenberg D, Von Kuster G, Bouvier E, Baker D, Afgan E et al. Dissemination of scientific software with galaxy toolShed. Genome Biol 2014; 15:403 [View Article] [PubMed]
    [Google Scholar]
  11. Miltgen G, Martak D, Valot B, Kamus L, Garrigos T et al. One health compartmental analysis of ESBL-producing Escherichia coli on reunion island reveals partitioning between humans and livestock. J Antimicrob Chemother 2022; 77:1254–1262 [View Article] [PubMed]
    [Google Scholar]
  12. Bouiller K, Zeggay A, Gbaguidi-Haore H, Hocquet D, Chirouze C et al. Epidemiology and risk factors of nasal carriage of Staphylococcus aureus CC398 in two distinct cohorts in France. Front Microbiol 2022; 13:1068420 [View Article] [PubMed]
    [Google Scholar]
  13. Martin M. Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet j 2011; 17:10–12 [View Article]
    [Google Scholar]
  14. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 2012; 19:455–477 [View Article] [PubMed]
    [Google Scholar]
  15. Li H. Aligning sequence reads, clone sequences and assembly Contigs with BWA-MEM. arXiv 2013 [View Article]
    [Google Scholar]
  16. Garrison E, Marth G. Haplotype-based variant detection from short-read sequencing. arXiv 2012 [View Article]
    [Google Scholar]
  17. Mellmann A, Bletz S, Böking T, Kipp F, Becker K et al. Real-time genome sequencing of resistant bacteria provides precision infection control in an institutional setting. J Clin Microbiol 2016; 54:2874–2881 [View Article] [PubMed]
    [Google Scholar]
  18. Palma F, Mangone I, Janowicz A, Moura A, Chiaverini A et al. In vitro and in silico parameters for precise cgMLST typing of Listeria monocytogenes. BMC Genomics 2022; 23:235 [View Article] [PubMed]
    [Google Scholar]
  19. Parcell BJ, Oravcova K, Pinheiro M, Holden MTG, Phillips G et al. Pseudomonas aeruginosa intensive care unit outbreak: winnowing of transmissions with molecular and genomic typing. J Hosp Infect 2018; 98:282–288 [View Article] [PubMed]
    [Google Scholar]
  20. Hammerum AM, Porsbo LJ, Hansen F, Roer L, Kaya H et al. Surveillance of OXA-244-producing Escherichia coli and epidemiologic investigation of cases, Denmark, January 2016 to August 2019. Euro Surveill 2020; 25:1900742 [View Article] [PubMed]
    [Google Scholar]
  21. Potron A, Hocquet D, Triponney P, Plésiat P, Bertrand X et al. Carbapenem-susceptible OXA-23-producing proteus mirabilis in the French community. Antimicrob Agents Chemother 2019; 63:e00191-19 [View Article] [PubMed]
    [Google Scholar]
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