Betül GİRAY, Füsun Bahriye UÇAR, Sabire Şöhret AYDEMİR

Genotypic analysis of Escherichia coli strains that cause urosepsis in the Aegean region

Background/aim: The aim of this study was to characterize strains genotypically, to determine their phylogenetic relationships, to investigate the presence of the papG gene, and to compare their antibiotic susceptibility test results. Materials and methods: Seventy pathogenic E. coli strains were isolated from both urine and blood cultures of patients with the preliminary diagnosis of urosepsis who were referred to the Ege University Faculty of Medicine, Bacteriology Laboratory of Medical Microbiology Department in İzmir. All of these strains were examined for the papG gene and phylogenetic groups with the multiplex polymerase chain reaction technique. Pulsed-field gel electrophoresis and multilocus sequence typing (MLST) were used for epidemiologic analysis. Results: Phylogenetically, it was found that 16 belonged to group B2, 31 belonged to group D, 15 belonged to group A, and 7 belonged to group B1. One strain was not identified as belonging to a group. papG genes were found in 26 of 70 E. coli strains. Thirty urosepsis pathogenic E. coli strains were analyzed with MLST. Twenty-two strains were identified as new STs. Conclusion: These findings are extremely important for Turkey and these new 22 strains should be investigated in more detail because they are new and have the potential to lead to infections

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1. Johnson JR, Russo AT. Molecular epidemiology of extraintestinal pathogenic (uropathogenic) Escherichia coli. Int J Med Microbiol 2005; 295: 383-404.
2. Palaniappan RU, Zhang Y, Chiu D, Torres A, Debroy C, Whittam TS, Chang YF. Differentiation of Escherichia coli pathotypes by oligonucleotide spotted array. J Clin Microbiol 2006; 44: 1495-1501.
3. Lloyd AL, Rasko DA, Mobley HL. Defining genomic islands and uropathogen-specific genes in uropathogenic Escherichia coli. J Bacteriol 2007; 189: 3532-3546.
4. Mora A, Lopez C, Dabhi G, Blanco M, Blanco JE, Alonso MP, Herrera A, Mamani R, Bonacorsi S, Molulin-Schouleur M et al. Extraintestinal pathogenic Escherichia coli O1:K1:H7/ NM from human and avian origin: detection of clonal groups B2 ST95 and D ST59 with different host distribution. BMC Microbiol 2009; 9: 1-11.
5. Kariyawasam S, Scaccianoce JA, Nolan LK. Common and specific genomic sequences of avian and human extraintestinal pathogenic Escherichia coli as determined by genomic subtractive hybridization. BMC Microbiol 2007; 7: 1-8.
6. Hacker J, Carnial E. Ecological fitness, genomics islands and bacterial pathogenicity. EMBO reports 2001; 376-381.
7. Oelschlaeger AT, Dobrindt U, Hacker J. Pathogenicity islands of uropathogenic E.coli and the evolution of virulence. Int J Antimicrob Agents 2002; 19: 517-521.
8. Boyd EF, Hartl DL. Chromosomal regions specific to pathogenic isolates of Escherichia coli have a phylogenetically clustered distribution. J Bacteriol 1998; 180: 1159-1165.
9. Clermont O, Bonacorsi S, Bingen E. Rapid and simple determination of the Escherichia coli phylogenetic group. App Environ Microbiol 2000; 66: 4555-4558.
10. Ramos LN, Saayman LM, Chapman AT, Tucker RJ, Smith HV, Faoagali J, Chin JC, Brauner A, Katouli M. Genetic relatedness and virulence gene profiles of Escherichia coli strains isolated from septicaemic and uroseptic patients. Eur J Clin Microbiol Infect Dis 2010; 29: 15-23.
11. Köhler CD, Dobrindt U. What defines extraintestinal pathogenic Escherichia coli? Int J Med Microbiol 2011; 301: 642-647.
12. Wagenlehrer FME, Weidner W, Naber KG, Optimal management of urosepsis from the urological perspective. Int J Antimicrob Agents 2007; 30: 390-397.
13. Wagenlehrer FME, Pilatz A, Naber KG. Therapeutic challenges of urosepsis. Eur J Clin Invest 2008; 38: 45-49.
14. Dhakal BK, Kulesus RR, Mulvey AM. Mechanisms and consequences of bladder cell invasion by uropathogenic Escherichia coli. Eur J Clin Invest Rev 2008; 38: 2-11.
15. Mamıkoğlu L, İnan D. İdrar Yolu Enfeskiyonları, Topçu WA, Söyletir G, Doğanay M. Enfeksiyon Hastalıkları ve Mikrobiyolojisi. 3rd ed, vol 2. İstanbul, Turkey: Nobel Tıp Kitabevleri; 2008. pp. 1487-1506.
16. Hagan EC, Mobley HL. Uropathogenic E. coli outer membrane antigens expressed during tract infection. Infect Immun 2007; 75: 3941-3949.
17. Martinez JA, Soto S, Fabrega A, Almela M, Mensa J, Sariano A, Marco F, Jimenez de Anta MT, Vila J. Relationship of phylogenetic background, biofilm production, and time to detection of growth in blood culture vials with clinical variables and prognosis associated with Escherichia coli bacteremia. J Clin Microbiol 2006; 44: 1468-1474.
18. Liu D, Coloe S, Baird R. Rapid mini preparation of fungal DNA for PCR. J Clin Microbiol 2000; 38: 471.
19. Giray B, Uçar FB, Aydemir Ş. Characterization of uropathogenic Escherichia coli strains obtained from urology outpatient clinic of Ege Medical Faculty in İzmir. Turk J Med Sci 2012; 42: 1328- 1337.
20. Johnson JR, Stell AL. Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise. J Infect Dis 2000; 181: 261- 72.
21. Durmaz R, Otlu B, Çalışkan A, Gürsoy A. Acinetobacter baumannii, Escherichia coli ve Klebsiella Türlerinin Moleküler Tiplendirilmesinde Kullanilabilecek Kisa Süreli pulsed-field gel elektroforez (PFGE) protokolü. Ankem Derg 2007; 21: 113-117.
22. Turabelidze D, Kotetishvili M, Kreger A, Morris JG Jr, Sulakvelidze A. Improved pulsed-field gel electrophoresis for typing vancomycin-resistant enterococci. J Clin Microbiol 2000; 38: 4242-4245.
23. Herzer PJ, Inouye M, Whitmann TS. Phylogenetic distribution of branched RNA-linked multicopy single-stranded DNA among natural isolates of Escherichia coli. J Bacteriol 1990; 172: 6175-6181.
24. Pitout JD. Extraintestinal pathogenic Escherichia coli: a combination of virulence with antibiotic resistance. Front Microbiol 2012; 3: 1-7.
25. Mokady D, Gophna U, Ro EZ. Extensive gene diversity in septisemic Escherichia coli strains. J Clin Microbiol 2005; 43: 66-73.
26. Mao BH, Chang YF, Scaria J, Chang C, Chou LW, Tien N, Wu, JJ, Tseng CC, Wang MC, Chang CC et al. Identification of Escherichia coli genes associated with urinary tract infections. J Clin Microbiol 2011; 50: 449-456.
27. Rasmussen LS, Ejrnaes K, Lundgren B, Hammerum AM, Moller NF. Virulence factors and phylogenetic grouping of Escherichia coli isolates from patients with bacteremia of urinary tract origin relate to sex and hospital- vs. communityacquired origin. Int J Med Microbiol 2012; 302: 129-134.
28. Curova K, Kmetova M, Vargova R, Lovayova V, Siegfried L. Toxins of Escherichia coli isolated from blood culture. Clin Microbiol 2014; 3: 1-5.
29. Johnson JR, Russo TA. Molecular epidemiology of extraintestinal pathogenic (uropathogenic) Escherichia coli. Int J Med Microbiol Rev 2005; 295: 383-404.
30. Ewers C, Li C, Wilking H, Kiesling S, Alt K, Antao EM, Laturns C, Diehl I, Glodde S, Homeir T et al. Avian pathogenic, uropathogenic, and newborn meningitis-causing Escherichia coli: how closely related are they? IJMM 2007; 297: 163-176.
31. Yumuk Z, Afacan G, Nicolas-Chanoine MH, Sotto A, Lavigne JP. Turkey: a further country concerned by communityacquired Escherichia coli clone O25-ST131 producing CTX-M-15. J Antimicrob Chemother 2008; 62: 284-288.
32. Koga VL, Tomazetto G, Cyoia PS, Neves MS, Vidotto MC, Nakazato G, Kobayashi RKT. Molecular screening of virulence genes in extraintestinal pathogenic Escherichia coli isolated from human blood culture in Brazil. Biomed Res Int 2014; 2014: 1-9.
33. Wiles JT, Kulesus RR, Mulvey AM. Origins and virulence mechanisms of uropathogenic Escherichia coli. Exp Mol Pathol 2008; 85: 11-19.
34. Bergsten G, Wullt B, Svanborg C. Escherichia coli, fimbriae, bacterial persistence and host response induction in the human urinary tract. Int J Med Microbiol 2005; 295: 487-502.
35. Johnson JR. papG Alleles among Escherichia coli strains causing urosepsis: associations with other bacterial characteristics and host comprise. Infect Immun 1998; 66: 4568-4571.
36. Moreno E, Planells I, Prats G, Planes AM, Moreno G, Andreu A. Comparative study of Escherichia coli virulence determinants in strains causing urinary tract bacteremia versus strains causing pyelonephritis and other sources of bacteremia. Diag Microbiol 2005; 53: 93-99.
37. Arısoy M, Rad AY, Akın A, Akar N. Relationship between susceptibility to antimicrobials and virulence factors in pediatric Escherichia coli isolates. Int J Antimicrob 2008; 31: 4-8.
38. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 1995; 33: 2233-2239.
39. Bidet P, Messai F, Blanco J, Blanco J, Dehem M, Aujard Y, Bingen E, Bonacorsi S. Combined multilocus sequence typing and O serogrouping distinguishes Escherichia coli subtypes associated with infant urosepsis and/or meningitis. JID 2007; 196: 297-303.
40. Tekeli A. Pulsed-field Gel Electrophoresis, Moleküler Mikrobiyoloji: Tanı Prensipleri ve Uygulamalar, Ankara, Turkey: Palme Yayıncılık; 2006.
41. Chanoine MH, Blanco J, Guibout VL, Demarty R, Alonso MP, Caniça MM, Park YJ, Lavigne JP, Pitout J, Johnson JR. Intercontinental emergence of Escherichia coli clone O25: H4- ST131 producing CTX-M-15. J Antimicrob Chemoter 2008; 61: 273-281.
42. Tartof SY, Solberg OW, Manges AR, Riley LW. Analysis of a uropathogenic Escherichia coli clonal group by multilocus sequence typing. J Clin Microbiol 2005; 43: 5860-5864.
43. Mora A, Dabhi G, Lopez C. Virulence patterns in murine sepsis model of ST131 Escherichia coli clinical isolates belonging to serotypes O25b:H4 and O16:H5 are associated to specific virotypes. Plos One 2014; 9: 1-9.
44. Anvarinejad M, Farshad SH, Ranjbar R, Giammanco GM, Alborzi A, Japoni A. Genotypic analysis of E. coli strains isolated from patient with cystitis and pyelonephritis. Iran Red Crescent Med J 2012; 14: 408-416.
45. Moulin-Schouleur M, Schouler C, Tailliez P, Kao MR, Bree A, Germon P, Oswald E, Mainil J, Blanco M, Blanco J. Common virulence factors and genetic relationships between O18:K1:H7 Escherichia coli isolates of human and avian origin. J Clin Microbiol 2006; 44: 3484-3492.
46. Usein CR, Grigore LA, Georgescu RM, Baltoiu MC, Condei M, Teleman MD. Phylogenetic background and extraintetsinal virulence genotypes of Escherichia coli vaginal isolated from adult women. Rev Romana Med Lab 2011; 19: 37-45.
47. Görgeç S, Kuzucu Ç, Otlu B, Yetkin F, Ersoy Y. Investigation of beta-lactamase genes and clonal relationship among the extended-spectrum beta-lactamase producing nosocomial Escherichia coli isolates. Mikrobiyol Bul 2015; 49: 15-25.
48. İnan NU, Gürler N. Investigation of antibiotic resistance and some virulence factors of Escherichia coli strains isolated from children with urinary tract infections. Ankem Derg 2004; 18: 89-96.
49. Vila J, Simon K, Ruiz J, Horcajada P, Velasco M, Barranco M, Moreno A, Mensa J. Are quinolone-resistant uropathogenic Escherichia coli less virulent? JID; 186: 1039-1042.
50. Johnson JR, Murray AC, Gajewski A. Isolation and molecular characterization of nalidixic acid-resistant extraintestinal pathogenic Escherichia coli from retail chicken products. Antimicrob Agents Chemother 2003; 47: 2161-2168.