1887

Abstract

Quorum sensing is known to regulate bacterial virulence, and the accessory gene regulator () loci is one of the genetic loci responsible for its regulation. Recent reports examining show that two loci, and regulate toxin production, but the diversity of loci and their epidemiology is unknown. In our study, analysis was performed to research genetic diversity of , and isolates from clinical samples underwent multilocus sequence typing (MLST) and PCR analysis of loci. To reveal the distribution of among different strains, phylogenetic analysis was also performed. In our analysis, two different subtypes, named and , were found in which were previously reported. PCR analysis of 133 . isolates showed that 131 strains had , 61 strains had , and 26 strains had was mainly found in clade 1 or clade 2 organisms, whereas was only found in clade 4. With rare exception, -negative sequence types (STs) belonged to clade C-Ⅰ and C-Ⅲ, and one clade 4 strain had . Our study revealed subtypes of not previously recognized, and the distribution of several loci in . These findings provide a foundation for further functional and clinical research of the loci.

Funding
This study was supported by the:
  • SHU OKUGAWA , JSPS , (Award 19K08949)
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/content/journal/acmi/10.1099/acmi.0.000134
2020-05-14
2020-06-04
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References

  1. Kuehne SA, Cartman ST, Heap JT, Kelly ML, Cockayne A et al. The role of toxin A and toxin B in Clostridium difficile infection. Nature 2010; 467:711–713 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  2. Bartlett JG, Chang TW, Gurwith M, Gorbach SL, Onderdonk AB. Antibiotic-associated pseudomembranous colitis due to toxin-producing clostridia. N Engl J Med 1978; 298:531–534 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  3. Whiteley M, Diggle SP, Greenberg EP. Progress in and promise of bacterial quorum sensing research. Nature 2017; 551:313–320 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  4. Ohtani K, Hayashi H, Shimizu T. The luxS gene is involved in cell-cell signalling for toxin production in Clostridium perfringens. Mol Microbiol 2002; 44:171–179 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  5. Sperandio V, Li CC, Kaper JB. Quorum-Sensing Escherichia coli regulator A: a regulator of the LysR family involved in the regulation of the locus of enterocyte effacement pathogenicity island in enterohemorrhagic E. coli. Infect Immun 2002; 70:3085–3093 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  6. Zhu J, Miller MB, Vance RE, Dziejman M, Bassler BL et al. Quorum-sensing regulators control virulence gene expression in Vibrio cholerae. Proc Natl Acad Sci U S A 2002; 99:3129–3134 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  7. Martin MJ, Clare S, Goulding D, Faulds-Pain A, Barquist L et al. The agr locus regulates virulence and colonization genes in Clostridium difficile 027. J Bacteriol 2013; 195:3672–3681 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  8. Darkoh C, DuPont HL, Norris SJ, Kaplan HB. Toxin synthesis by Clostridium difficile is regulated through quorum signaling. mBio 2015; 6:e02569 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  9. Darkoh C, Odo C, DuPont HL. Accessory gene Regulator-1 locus is essential for virulence and pathogenesis of Clostridium difficile. mBio 2016; 7:e01237-16 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  10. Wuster A, Babu MM. Conservation and evolutionary dynamics of the agr cell-to-cell communication system across Firmicutes. J Bacteriol 2008; 190:743–746 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  11. Qin X, Singh KV, Weinstock GM, Murray BE. Effects of Enterococcus faecalis fsr genes on production of gelatinase and a serine protease and virulence. Infect Immun 2000; 68:2579–2586 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  12. Autret N, Raynaud C, Dubail I, Berche P, Charbit A. Identification of the agr locus of Listeria monocytogenes: role in bacterial virulence. Infect Immun 2003; 71:4463–4471 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  13. Marsden GL, Davis IJ, Wright VJ, Sebaihia M, Kuijper EJ et al. Array comparative hybridisation reveals a high degree of similarity between UK and European clinical isolates of hypervirulent Clostridium difficile. BMC Genomics 2010; 11:389 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  14. Okada Y, Yagihara Y, Wakabayashi Y, Igawa G, Saito R et al. Epidemiology and virulence-associated genes of Clostridioides difficile isolates and factors associated with toxin EIA results at a university hospital in Japan. Access Microbiol 2020; 2: [CrossRef]
    [Google Scholar]
  15. Kuwata Y, Tanimoto S, Sawabe E, Shima M, Takahashi Y et al. Molecular epidemiology and antimicrobial susceptibility of Clostridium difficile isolated from a university teaching hospital in Japan. Eur J Clin Microbiol Infect Dis 2015; 34:763–772 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  16. Persson S, Torpdahl M, Olsen KEP. New multiplex PCR method for the detection of Clostridium difficile toxin A (tcdA) and toxin B (tcdB) and the binary toxin (cdtA/cdtB) genes applied to a Danish strain collection. Clin Microbiol Infect 2008; 14:1057–1064 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  17. Talevich E, Invergo BM, Cock PJA, Chapman BA. Bio.Phylo: a unified toolkit for processing, analyzing and visualizing phylogenetic trees in Biopython. BMC Bioinformatics 2012; 13:209 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  18. Cock PJA, Antao T, Chang JT, Chapman BA, Cox CJ et al. Biopython: freely available python tools for computational molecular biology and bioinformatics. Bioinformatics 2009; 25:1422–1423 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  19. Guindon S, Dufayard J-F, Lefort V, Anisimova M, Hordijk W et al. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 2010; 59:307–321 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  20. Ciccarelli FD, Doerks T, von Mering C, Creevey CJ, Snel B et al. Toward automatic reconstruction of a highly resolved tree of life. Science 2006; 311:1283–1287 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  21. Dingle KE, Elliott B, Robinson E, Griffiths D, Eyre DW et al. Evolutionary history of the Clostridium difficile pathogenicity locus. Genome Biol Evol 2014; 6:36–52 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  22. Chen R, Feng Y, Wang X, Yang J, Zhang X et al. Whole genome sequences of three clade 3 Clostridium difficile strains carrying binary toxin genes in China. Sci Rep 2017; 7:43555 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  23. Kim J, Kim Y, Pai H. Clinical characteristics and treatment outcomes of Clostridium difficile infections by PCR ribotype 017 and 018 strains. PLoS One 2016; 11:e0168849 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  24. Knight DR, Elliott B, Chang BJ, Perkins TT, Riley TV. Diversity and evolution in the genome of Clostridium difficile. Clin Microbiol Rev 2015; 28:721–741 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  25. Ramírez-Vargas G, López-Ureña D, Badilla A, Orozco-Aguilar J, Murillo T et al. Novel clade C-I Clostridium difficile strains escape diagnostic tests, differ in pathogenicity potential and carry toxins on extrachromosomal elements. Sci Rep 2018; 8:13951 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  26. Imwattana K, Knight DR, Kullin B, Collins DA, Putsathit P et al. Clostridium difficile ribotype 017 - characterization, evolution and epidemiology of the dominant strain in Asia. Emerg Microbes Infect 2019; 8:796–807 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  27. Stabler RA, He M, Dawson L, Martin M, Valiente E et al. Comparative genome and phenotypic analysis of Clostridium difficile 027 strains provides insight into the evolution of a hypervirulent bacterium. Genome Biol 2009; 10:R102 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  28. Riedel T, Wetzel D, Hofmann JD, Plorin SPEO, Dannheim H et al. High metabolic versatility of different toxigenic and non-toxigenic Clostridioides difficile isolates. Int J Med Microbiol 2017; 307:311–320 [CrossRef][PubMed][PubMed]
    [Google Scholar]
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