1887
Research article Open Access
Like 0

Abstract

Introduction

At the beginning of 2016, an increase in paediatric haemolytic uremic syndrome (HUS) cases was observed in Romania. The microbiological investigations allowed isolation of Shiga toxin-producing (STEC) O26 as the causative agent from most cases. An enhanced national surveillance of HUS and severe diarrhoea was established across the country following the identification of the first cases and was carried out until August 2016. A total of 15 strains were isolated from 10 HUS and five diarrhoea cases. Strains were characterised by virulence markers (i.e. type/subtype, , genes), phylogroup, genetic relatedness and clonality using PCR-based assays, PFGE and multilocus sequence typing (MLST). The first six strains were further characterised by whole genome sequencing (WGS). Five PCR-defined genotypes were distinguished. All strains from HUS cases harboured and , with or without , while strains from diarrhoea cases carried exclusively and genes. PFGE resolved strains into multiple pulsotypes, compatible with a certain geographic segregation of the cases, and strains were assigned to phylogroup B1 and sequence type (ST) 21. WGS confirmed the results of conventional molecular methods, brought evidence of O26:H11 serotype, and complemented the virulence profiles. This first description of STEC O26 strains from cases in Romania showed that the isolates belonged to a diverse population. The virulence content of most strains highlighted a high risk for severe outcome in infected patients. Improving the national surveillance strategy for STEC infections in Romania needs to be further considered.

Loading

Article metrics loading...

/content/10.2807/1560-7917.ES.2017.22.47.17-00148
2017-11-23
2017-12-11
http://instance.metastore.ingenta.com/content/10.2807/1560-7917.ES.2017.22.47.17-00148
Loading
Loading full text...

Full text loading...

/deliver/fulltext/eurosurveillance/22/47/eurosurv-22-47-3.html?itemId=/content/10.2807/1560-7917.ES.2017.22.47.17-00148&mimeType=html&fmt=ahah

References

  1. Caprioli A, Morabito S, Brugère H, Oswald E. Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission. Vet Res. 2005;36(3):289-311.  https://doi.org/10.1051/vetres:2005002  PMID: 15845227 
  2. Adams NL, Byrne L, Smith GA, Elson R, Harris JP, Salmon R, et al. Shiga Toxin-Producing Escherichia coli O157, England and Wales, 1983-2012. Emerg Infect Dis. 2016;22(4):590-7.  https://doi.org/10.3201/eid2204.151485  PMID: 26982243 
  3. Brandal LT, Wester AL, Lange H, Løbersli I, Lindstedt BA, Vold L, et al. Shiga toxin-producing escherichia coli infections in Norway, 1992-2012: characterization of isolates and identification of risk factors for haemolytic uremic syndrome. BMC Infect Dis. 2015;15(1):324.  https://doi.org/10.1186/s12879-015-1017-6  PMID: 26259588 
  4. Heiman KE, Mody RK, Johnson SD, Griffin PM, Gould LH. Escherichia coli O157 Outbreaks in the United States, 2003-2012. Emerg Infect Dis. 2015;21(8):1293-301.  https://doi.org/10.3201/eid2108.141364  PMID: 26197993 
  5. Tarr PI, Gordon CA, Chandler WL. Shiga-toxin-producing Escherichia coli and haemolytic uraemic syndrome. Lancet. 2005;365(9464):1073-86.  https://doi.org/10.1016/S0140-6736(05)71144-2  PMID: 15781103 
  6. Germinario C, Caprioli A, Giordano M, Chironna M, Gallone MS, Tafuri S, et al. Community-wide outbreak of haemolytic uraemic syndrome associated with Shiga toxin 2-producing Escherichia coli O26:H11 in southern Italy, summer 2013. Euro Surveill. 2016;21(38):30343.  https://doi.org/10.2807/1560-7917.ES.2016.21.38.30343  PMID: 27684204 
  7. Kuehne A, Bouwknegt M, Havelaar A, Gilsdorf A, Hoyer P, Stark K, et al. Estimating true incidence of O157 and non-O157 Shiga toxin-producing Escherichia coli illness in Germany based on notification data of haemolytic uraemic syndrome. Epidemiol Infect. 2016;144(15):3305-15.  https://doi.org/10.1017/S0950268816001436  PMID: 27468812 
  8. Luna-Gierke RE, Griffin PM, Gould LH, Herman K, Bopp CA, Strockbine N, et al. Outbreaks of non-O157 Shiga toxin-producing Escherichia coli infection: USA. Epidemiol Infect. 2014;142(11):2270-80.  https://doi.org/10.1017/S0950268813003233  PMID: 24398154 
  9. Messens W, Bolton D, Frankel G, Liebana E, McLauchlin J, Morabito S, et al. Defining pathogenic verocytotoxin-producing Escherichia coli (VTEC) from cases of human infection in the European Union, 2007-2010. Epidemiol Infect. 2015;143(8):1652-61.  PMID: 25921781 
  10. Dallman TJ, Byrne L, Launders N, Glen K, Grant KA, Jenkins C. The utility and public health implications of PCR and whole genome sequencing for the detection and investigation of an outbreak of Shiga toxin-producing Escherichia coli serogroup O26:H11. Epidemiol Infect. 2015;143(8):1672-80.  https://doi.org/10.1017/S0950268814002696  PMID: 25316375 
  11. Bielaszewska M, Mellmann A, Bletz S, Zhang W, Köck R, Kossow A, et al. Enterohemorrhagic Escherichia coli O26:H11/H-: a new virulent clone emerges in Europe. Clin Infect Dis. 2013;56(10):1373-81.  https://doi.org/10.1093/cid/cit055  PMID: 23378282 
  12. Peron E, Zaharia A, Zota LC, Severi E, Mårdh O, Usein C, et al. Early findings in outbreak of haemolytic uraemic syndrome among young children caused by Shiga toxin-producing Escherichia coli, Romania, January to February 2016. Euro Surveill. 2016;21(11):30170.  https://doi.org/10.2807/1560-7917.ES.2016.21.11.30170  PMID: 27020906 
  13. European Centre for Disease Prevention and Control (ECDC) and European Food Safety Authority. (EFSA). Multi-country outbreak of Shiga toxin-producing Escherichia coli infection associated with haemolytic uraemic syndrome. Stockholm: ECDC; 5 Apr 2016. Available from: http://ecdc.europa.eu/en/publications/Publications/RRA-Escherichia-coli-O26-Romania-Italy-April2016.pdf
  14. Schmidt H, Beutin L, Karch H. Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933. Infect Immun. 1995;63(3):1055-61. PMID: 7868227 
  15. Scheutz F, Teel LD, Beutin L, Piérard D, Buvens G, Karch H, et al. Multicenter evaluation of a sequence-based protocol for subtyping Shiga toxins and standardizing Stx nomenclature. J Clin Microbiol. 2012;50(9):2951-63.  https://doi.org/10.1128/JCM.00860-12  PMID: 22760050 
  16. Clermont O, Christenson JK, Denamur E, Gordon DM. The Clermont Escherichia coli phylo-typing method revisited: improvement of specificity and detection of new phylo-groups. Environ Microbiol Rep. 2013;5(1):58-65.  https://doi.org/10.1111/1758-2229.12019  PMID: 23757131 
  17. Ribot EM, Fair MA, Gautom R, Cameron DN, Hunter SB, Swaminathan B, et al. Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne Pathog Dis. 2006;3(1):59-67.  https://doi.org/10.1089/fpd.2006.3.59  PMID: 16602980 
  18. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995;33(9):2233-9. PMID: 7494007 
  19. Wirth T, Falush D, Lan R, Colles F, Mensa P, Wieler LH, et al. Sex and virulence in Escherichia coli: an evolutionary perspective. Mol Microbiol. 2006;60(5):1136-51.  https://doi.org/10.1111/j.1365-2958.2006.05172.x  PMID: 16689791 
  20. European Union Reference Laboratory VTEC. Advanced Research Infrastructure for Experimentation in Genomics (ARIES) public Galaxy server. Rome: Istituto Superiore di Sanità (ISS). [Accessed 20 Jun 2016]. Available from: https://w3.iss.it/site/aries/
  21. Joensen KG, Scheutz F, Lund O, Hasman H, Kaas RS, Nielsen EM, et al. Real-time whole-genome sequencing for routine typing, surveillance, and outbreak detection of verotoxigenic Escherichia coli. J Clin Microbiol. 2014;52(5):1501-10.  https://doi.org/10.1128/JCM.03617-13  PMID: 24574290 
  22. Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nat Methods. 2012;9(4):357-9.  https://doi.org/10.1038/nmeth.1923  PMID: 22388286 
  23. 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(5):455-77.  https://doi.org/10.1089/cmb.2012.0021  PMID: 22506599 
  24. 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(8):2410-26.  https://doi.org/10.1128/JCM.00008-15  PMID: 25972421 
  25. Inouye M, Dashnow H, Raven LA, Schultz MB, Pope BJ, Tomita T, et al. SRST2: Rapid genomic surveillance for public health and hospital microbiology labs. Genome Med. 2014;6(11):90.  https://doi.org/10.1186/s13073-014-0090-6  PMID: 25422674 
  26. Gardner SN, Slezak T, Hall BG. kSNP3.0: SNP detection and phylogenetic analysis of genomes without genome alignment or reference genome. Bioinformatics. 2015;31(17):2877-8.  https://doi.org/10.1093/bioinformatics/btv271  PMID: 25913206 
  27. Wilks SS. The Large-Sample Distribution of the Likelihood Ratio for Testing Composite Hypotheses. Ann Math Stat. 1938;9(1):60-2.  https://doi.org/10.1214/aoms/1177732360 
  28. 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(3):497-503. PMID: 9986802 
  29. Persson S, Olsen KE, Ethelberg S, Scheutz F. Subtyping method for Escherichia coli shiga toxin (verocytotoxin) 2 variants and correlations to clinical manifestations. J Clin Microbiol. 2007;45(6):2020-4.  https://doi.org/10.1128/JCM.02591-06  PMID: 17446326 
  30. Werber D, Fruth A, Buchholz U, Prager R, Kramer MH, Ammon A, et al. Strong association between shiga toxin-producing Escherichia coli O157 and virulence genes stx2 and eae as possible explanation for predominance of serogroup O157 in patients with haemolytic uraemic syndrome. Eur J Clin Microbiol Infect Dis. 2003;22(12):726-30.  https://doi.org/10.1007/s10096-003-1025-0  PMID: 14614596 
  31. Buvens G, De Gheldre Y, Dediste A, de Moreau AI, Mascart G, Simon A, et al. Incidence and virulence determinants of verocytotoxin-producing Escherichia coli infections in the Brussels-Capital Region, Belgium, in 2008-2010. J Clin Microbiol. 2012;50(4):1336-45.  https://doi.org/10.1128/JCM.05317-11  PMID: 22238434 
  32. Kaper JB, Nataro JP, Mobley HL. Pathogenic Escherichia coli. Nat Rev Microbiol. 2004;2(2):123-40.  https://doi.org/10.1038/nrmicro818  PMID: 15040260 
  33. Brunder W, Schmidt H, Frosch M, Karch H. The large plasmids of Shiga-toxin-producing Escherichia coli (STEC) are highly variable genetic elements. Microbiology. 1999;145(5):1005-14.  https://doi.org/10.1099/13500872-145-5-1005  PMID: 10376815 
  34. Fratamico PM, Yan X, Caprioli A, Esposito G, Needleman DS, Pepe T, et al. The complete DNA sequence and analysis of the virulence plasmid and of five additional plasmids carried by Shiga toxin-producing Escherichia coli O26:H11 strain H30. Int J Med Microbiol. 2011;301(3):192-203.  https://doi.org/10.1016/j.ijmm.2010.09.002  PMID: 21212019 
  35. Marejková M, Bláhová K, Janda J, Fruth A, Petráš P. Enterohemorrhagic Escherichia coli as causes of hemolytic uremic syndrome in the Czech Republic. PLoS One. 2013;8(9):e73927.  https://doi.org/10.1371/journal.pone.0073927  PMID: 24040117 
  36. Januszkiewicz A, Wołkowicz T, Chróst A, Szych J. Characterization of the Shiga toxin-producing Escherichia coli O26 isolated from human in Poland between 1996 and 2014. Lett Appl Microbiol. 2015;60(6):605-8.  https://doi.org/10.1111/lam.12413  PMID: 25758912 
/content/10.2807/1560-7917.ES.2017.22.47.17-00148
Loading

Data & Media loading...

Comment has been disabled for this content
Submit comment
Close
Comment moderation successfully completed
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error