Skip to main content

The CGE Tool Box

  • Chapter
  • First Online:
Applied Genomics of Foodborne Pathogens

Abstract

As whole genome sequence data of microorganisms are becoming easily accessible and cheap to produce, a transformation of the traditional methods used for typing, phenotyping and phylogenetic analysis of microorganisms is on the way. Following the anticipation that most clinical microbiological and food safety laboratories will soon have a sequencer in use on a daily basis, there is a growing need for easy-to-use bioinformatics methods that can quickly convert the sequence data into useful information on, e.g., the type of bacteria, whether it is resistant towards any types of antibiotics, and whether it is part of an outbreak. The Center for Genomic Epidemiology, which is located at the Technical University of Denmark, has since its beginning in 2010 developed such bioinformatics methods and made them freely available as web-services. These web-services and their use is the focus of this chapter.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 119.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Notes

  1. 1.

    VCF (Variant Call Format) files specify a type of text files used for storing sequence variation.

References

  1. Aarestrup FM, Brown EW, Detter C, Gerner-Smidt P, Gilmour D, Harmsen MW, Hendriksen RS, Hewson R, Heymann DL, Johansson K, Ijaz K, Keim PS, Koopmans M, Kroneman A, Lo Fo Wong D, Lund O, Palm D, Sawanpanyalert P, Sobel J, Schlundt J. Integrating genome-based informatics to modernize global disease monitoring, information sharing, and response. Emerg Infect Dis. 2012;18, e1.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Kaas RS, Leekitcharoenphon P, Aarestrup FM, Lund O. Solving the problem of comparing whole bacterial genomes across different sequencing platforms. PLoS One. 2014;9, e104984.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Hasman H, Saputra D, Sicheritz-Ponten T, Lund O, Svendsen CA, Frimodt-Moller N, Aarestrup FM. Rapid whole-genome sequencing for detection and characterization of microorganisms directly from clinical samples. J Clin Microbiol. 2014;52:139–46.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Larsen MV, Cosentino S, Rasmussen S, Friis C, Hasman H, Marvig RL, Jelsbak L, Sicheritz-Ponten T, Ussery DW, Aarestrup FM, Lund O. Multilocus sequence typing of total-genome-sequenced bacteria. J Clin Microbiol. 2012;50:1355–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Joensen KG, Scheutz F, Lund O, Hasman H, Kaas RS, Nielsen EM, Aarestrup FM. Real-time whole-genome sequencing for routine typing, surveillance, and outbreak detection of verotoxigenic Escherichia coli. J Clin Microbiol. 2014;52:1501–10.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Lukjancenko O, Thomsen MCF, Larsen MV, Ussery DW. PanFunPro: PAN-genome analysis based on FUNctional PROfiles [v1; ref status: approved with reservations 3]. F1000Research. 2013;2:265. http://f1000r.es/2e1.

    Google Scholar 

  7. Cosentino S, Voldby Larsen M, Moller Aarestrup F, Lund O. PathogenFinder—distinguishing friend from foe using bacterial whole genome sequence data. PLoS One. 2013;8, e77302.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Carattoli A, Zankari E, Garcia-Fernandez A, Voldby Larsen M, Lund O, Villa L, Moller Aarestrup F, Hasman H. In silico detection and typing of plasmids using PlasmidFinder and plasmid multilocus sequence typing. Antimicrob Agents Chemother. 2014;58:3895–903.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Zankari E, Hasman H, Cosentino S, Vestergaard M, Rasmussen S, Lund O, Aarestrup FM, Larsen MV. Identification of acquired antimicrobial resistance genes. J Antimicrob Chemother. 2012;67:2640–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Joensen KG, Tetzschner AM, Iguchi A, Aarestrup FM, Scheutz F. Rapid and easy in silico serotyping of Escherichia coli isolates by use of whole-genome sequencing data. J Clin Microbiol. 2015;53:2410–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Leekitcharoenphon P, Mortensen RK, Thomsen MCF, Friis C, Rasmussen S, Aarestrup FM. snpTree—a web-server to identify and construct SNP trees from whole genome sequence data. BMC Genomics. 2012;13 Suppl 7:S6.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Larsen J, Enright MC, Godoy D, Spratt BG, Larsen AR, Skov RL. Multilocus sequence typing scheme for Staphylococcus aureus: revision of the gmk locus. J Clin Microbiol. 2012;50:2538–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Boerlin P, McEwen SA, Boerlin-Petzold F, Wilson JB, Johnson RP, Gyles CL. Associations between virulence factors of Shiga toxin-producing Escherichia coli and disease in humans. J Clin Microbiol. 1999;37:497–503.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Karmali MA. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989;2:15–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Karch H, Tarr PI, Bielaszewska M. Enterohaemorrhagic Escherichia coli in human medicine. Int J Med Microbiol. 2005;295:405–18.

    Article  CAS  PubMed  Google Scholar 

  16. CDC. Centers for Disease Control and Prevention—E. coli. General Information. 2012. http://www.cdc.gov/ecoli/general/#complications.

  17. Richter M, Rossello-Mora R. Shifting the genomic gold standard for the prokaryotic species definition. Proc Natl Acad Sci U S A. 2009;106:19126–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Fox GE, Peckman KJ, Woese CE. Comparative cataloging of 16S ribosomal ribonucleic acid: molecular approach to procaryotic systematics. Int J Syst Bacteriol. 1977;27:44–57.

    Article  CAS  Google Scholar 

  19. DeSantis TZ, Hugenholtz P, Larsen N, Rojas M, Brodie EL, Keller K, Huber T, Dalevi D, Hu P, Andersen GL. Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Appl Environ Microbiol. 2006;72:5069–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Pruesse E, Quast C, Knittel K, Fuchs BM, Ludwig W, Peplies J, Glockner FO. SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res. 2007;35:7188–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Kampfer P. Systematics of prokaryotes: the state of the art. Antonie Van Leeuwenhoek. 2012;101:3–11.

    Article  PubMed  Google Scholar 

  22. Tindall BJ, Rossello-Mora R, Busse HJ, Ludwig W, Kampfer P. Notes on the characterization of prokaryote strains for taxonomic purposes. Int J Syst Evol Microbiol. 2010;60:249–66.

    Article  CAS  PubMed  Google Scholar 

  23. Tindall BJ, Schneider S, Lapidus A, Copeland A, Glavina Del Rio T, Nolan M, Lucas S, Chen F, Tice H, Cheng JF, Saunders E, Bruce D, Goodwin L, Pitluck S, Mikhailova N, Pati A, Ivanova N, Mavrommatis K, Chen A, Palaniappan K, Chain P, Land M, Hauser L, Chang YJ, Jeffries CD, Brettin T, Han C, Rohde M, Goker M, Bristow J, Eisen JA, Markowitz V, Hugenholtz P, Klenk HP, Kyrpides NC, Detter JC. Complete genome sequence of Halomicrobium mukohataei type strain (arg-2). Stand Genomic Sci. 2009;1:270–7.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Klenk HP, Goker M. En route to a genome-based classification of Archaea and Bacteria? Syst Appl Microbiol. 2010;33:175–82.

    Article  CAS  PubMed  Google Scholar 

  25. Larsen MV, Cosentino S, Lukjancenko O, Saputra D, Rasmussen S, Hasman H, Sicheritz-Ponten T, Aarestrup FM, Ussery DW, Lund O. Benchmarking of methods for genomic taxonomy. J Clin Microbiol. 2014;52(5):1529–39.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Lukjancenko O, Ussery DW. Vibrio chromosome-specific families. Front Microbiol. 2014;5:73.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Goujon M, McWilliam H, Li W, Valentin F, Squizzato S, Paern J, Lopez R. A new bioinformatics analysis tools framework at EMBL-EBI. Nucleic Acids Res. 2010;38:W695–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Woese CR, Fox GE. Phylogenetic structure of the prokaryotic domain: the primary kingdoms. Proc Natl Acad Sci U S A. 1977;74:5088–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Helgason E, Okstad OA, Caugant DA, Johansen HA, Fouet A, Mock M, Hegna I, Kolsto AB. Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis—one species on the basis of genetic evidence. Appl Environ Microbiol. 2000;66:2627–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Karaolis DK, Lan R, Reeves PR. Sequence variation in Shigella sonnei (Sonnei), a pathogenic clone of Escherichia coli, over four continents and 41 years. J Clin Microbiol. 1994;32:796–802.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Lan R, Reeves PR. Escherichia coli in disguise: molecular origins of Shigella. Microbes Infect. 2002;4:1125–32.

    Article  CAS  PubMed  Google Scholar 

  32. Maiden MC, Bygraves JA, Feil E, Morelli G, Russell JE, Urwin R, Zhang Q, Zhou J, Zurth K, Caugant DA, Feavers IM, Achtman M, Spratt BG. Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc Natl Acad Sci U S A. 1998;95:3140–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Jaureguy F, Landraud L, Passet V, Diancourt L, Frapy E, Guigon G, Carbonnelle E, Lortholary O, Clermont O, Denamur E, Picard B, Nassif X, Brisse S. Phylogenetic and genomic diversity of human bacteremic Escherichia coli strains. BMC Genomics. 2008;9:560.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Wirth T, Falush D, Lan R, Colles F, Mensa P, Wieler LH, Karch H, Reeves PR, Maiden MC, Ochman H, Achtman M. Sex and virulence in Escherichia coli: an evolutionary perspective. Mol Microbiol. 2006;60:1136–51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Griffiths D, Fawley W, Kachrimanidou M, Bowden R, Crook DW, Fung R, Golubchik T, Harding RM, Jeffery KJ, Jolley KA, Kirton R, Peto TE, Rees G, Stoesser N, Vaughan A, Walker AS, Young BC, Wilcox M, Dingle KE. Multilocus sequence typing of Clostridium difficile. J Clin Microbiol. 2010;48:770–8.

    Article  CAS  PubMed  Google Scholar 

  36. Lemee L, Dhalluin A, Pestel-Caron M, Lemeland JF, Pons JL. Multilocus sequence typing analysis of human and animal Clostridium difficile isolates of various toxigenic types. J Clin Microbiol. 2004;42:2609–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997;25:3389–402.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Hendriksen RS, Joensen KG, Lukwesa-Musyani C, Kalondaa A, Leekitcharoenphon P, Nakazwe R, Aarestrup FM, Hasman H, Mwansa JC. Extremely drug-resistant Salmonella enterica serovar Senftenberg infections in patients in Zambia. J Clin Microbiol. 2013;51:284–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Rodriguez-Rivera LD, Moreno Switt AI, Degoricija L, Fang R, Cummings CA, Furtado MR, Wiedmann M, den Bakker HC. Genomic characterization of Salmonella Cerro ST367, an emerging Salmonella subtype in cattle in the United States. BMC Genomics. 2014;15:427.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Stegger M, Aziz M, Chroboczek T, Price LB, Ronco T, Kiil K, Skov RL, Laurent F, Andersen PS. Genome analysis of Staphylococcus aureus ST291, a double locus variant of ST398, reveals a distinct genetic lineage. PLoS One. 2013;8, e63008.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Zhang S, Yin Y, Jones MB, Zhang Z, Deatherage Kaiser BL, Dinsmore BA, Fitzgerald C, Fields PI, Deng X. Salmonella serotype determination utilizing high-throughput genome sequencing data. J Clin Microbiol. 2015;53:1685–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ. Identification of plasmids by PCR-based replicon typing. J Microbiol Methods. 2005;63:219–28.

    Article  CAS  PubMed  Google Scholar 

  43. Zankari E, Hasman H, Kaas RS, Seyfarth AM, Agerso Y, Lund O, Larsen MV, Aarestrup FM. Genotyping using whole-genome sequencing is a realistic alternative to surveillance based on phenotypic antimicrobial susceptibility testing. J Antimicrob Chemother. 2013;68:771–7.

    Article  CAS  PubMed  Google Scholar 

  44. Villa L, Feudi C, Fortini D, Garcia-Fernandez A, Carattoli A. Genomics of KPC-producing Klebsiella pneumoniae sequence type 512 clone highlights the role of RamR and ribosomal S10 protein mutations in conferring tigecycline resistance. Antimicrob Agents Chemother. 2014;58:1707–12.

    Article  PubMed  PubMed Central  Google Scholar 

  45. Leonard SR, Lacher DW, Elkins CA, Jung CM. Draft genome sequence of the multidrug-resistant Escherichia coli strain LR09, isolated from a wastewater treatment plant. Genome Announc. 2014;2, e00272-14.

    Article  PubMed  PubMed Central  Google Scholar 

  46. Kroft BS, Brown EW, Meng J, Gonzalez-Escalona N. Draft genome sequences of two Salmonella strains from the SARA collection, SARA64 (Muenchen) and SARA33 (Heidelberg). Provide insight into their antibiotic resistance. Genome Announc. 2013;1, e00806-13.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Andreatta M, Nielsen M, Moller Aarestrup F, Lund O. In silico prediction of human pathogenicity in the gamma-proteobacteria. PLoS One. 2010;5, e13680.

    Article  PubMed  PubMed Central  Google Scholar 

  48. Delcher AL, Phillippy A, Carlton J, Salzberg SL. Fast algorithms for large-scale genome alignment and comparison. Nucleic Acids Res. 2002;30:2478–83.

    Article  PubMed  PubMed Central  Google Scholar 

  49. Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009;25:1754–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, Marth G, Abecasis G, Durbin R. The sequence alignment/map format and SAM tools. Bioinformatics. 2009;25:2078–9.

    Article  PubMed  PubMed Central  Google Scholar 

  51. Guio H, Tarazona D, Galarza M, Borda V, Curitomay R. Genome analysis of 17 extensively drug-resistant strains reveals new potential mutations for resistance. Genome Announc. 2014;2, e00759-14.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Teo J, Tan SY, Liu Y, Tay M, Ding Y, Li Y, Kjelleberg S, Givskov M, Lin RT, Yang L. Comparative genomic analysis of malaria mosquito vector-associated novel pathogen Elizabethkingia anophelis. Genome Biol Evol. 2014;6:1158–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Soborg B, Lassen SG, Muller L, Jensen T, Ethelberg S, Molbak K, Scheutz F. A verocytotoxin-producing E. coli outbreak with a surprisingly high risk of haemolytic uraemic syndrome, Denmark, September–October 2012. Euro Surveill. 2013;18:1–3.

    Google Scholar 

  54. Nordahl Petersen T, Rasmussen S, Hasman H, Caroe C, Baelum J, Schultz AC, Bergmark L, Svendsen CA, Lund O, Sicheritz-Ponten T, Aarestrup FM. Meta-genomic analysis of toilet waste from long distance flights; a step towards global surveillance of infectious diseases and antimicrobial resistance. Sci Rep. 2015;5:11444.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The Center for Genomic Epidemiology at the Technical University of Denmark is funded by grant 09-067103/DSF from the Danish Council for Strategic Research.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Mette Voldby Larsen .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer International Publishing Switzerland

About this chapter

Cite this chapter

Larsen, M.V. et al. (2017). The CGE Tool Box. In: Deng, X., den Bakker, H., Hendriksen, R. (eds) Applied Genomics of Foodborne Pathogens. Food Microbiology and Food Safety(). Springer, Cham. https://doi.org/10.1007/978-3-319-43751-4_5

Download citation

Publish with us

Policies and ethics