Molecular Identification and Antibiotic Resistance of Shigatoxigenic Escherichia Coli Strains in Children of the Age of Under 5-Years with Diarrhea in Yasuj, Iran

Document Type : Original Article(s)

Authors

1 Associate Professor, Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran

2 Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran

3 Assistant Professor, Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran

4 Young Researchers and Elite Club, Islamic Azad University, Marvdasht Branch, Marvdasht, Iran

Abstract

Background: Shigatoxigenic Escherichia coli (E. coli) strain is one of the most important causes of diarrhea, hemorrhagic colitis, and haemolytic uremic syndrome. The aim of this study was to survey the prevalence of shigatoxigenic strains and virulence genes in children of the age of under 5-years in Yasuj, Iran.Methods: This cross-sectional study was performed on 300 stool samples taken from children of the age of under 5-years with diarrhea in Yasuj. After initial identification of E. coli strains by culture and biochemical tests, shiga toxigenic Escherichia coli (STEC) genes such as eaeA, stx1 and stx2 detected by multiplex polymerase chain reaction (PCR) technique and antibiotic susceptibility of isolates was evaluated using disc diffusion method under the guidelines of Clinical and Laboratory Standards Institute (CLSI).Findings: Out of the samples, 104 (34.7%) STEC genes were separated. Out of considered virulence genes, 14 cases (13.46%) had stx1, 31 cases (29.81%)stx2, 9 cases (8.65%) genotype stx1-stx2, 12 cases (11.54%) eaeA-stx1, 25 cases (24.04%) eaeA-stx2 and 13 cases (12.5%) had third eaeA-stx1-stx2 genes. Antibiotic susceptibility test showed that the most susceptible antibiotic was imipenem for STEC; the most resistant antibiotic was ceftizoxime.Conclusion: Results showed that STEC strains have high prevalence in our study area. Therefore, hospital-wide surveillance using molecular techniques should be proposed in other regions of our country.

Keywords

References

  1. Reyes D, Vilchez S, Paniagua M, Colque-Navarro P, Weintraub A, Mollby R, et al. Diarrheagenic Escherichia coli markers and phenotypes among fecal E. coli isolates collected from Nicaraguan infants. J Clin Microbiol 2010; 48(9): 3395-6.
  2. Schultsz C, van den Ende J, Cobelens F, Vervoort T, van GA, Wetsteyn JC, et al. Diarrheagenic Escherichia coli and acute and persistent diarrhea in returned travelers. J Clin Microbiol 2000; 38(10): 3550-4.
  3. Nataro JP, Kaper JB. Diarrheagenic Escherichia coli. Clin Microbiol Rev 1998; 11(1): 142-201.
  4. Chang WS, Afsah-Hejri L, Rukayadi Y, Khatib A, Lye YL, Loo YY, et al. Quantification of Escherichia coli O157:H7 in organic vegetables and chickens. Int Food Res J 2013; 20(2): 1023-9.
  5. Kargar M, Daneshvar M, Homayoon M. Surveillance of virulence markers and antibiotic resistance of shiga toxin producing E.coli O157:H7 strains from meats purchase in Shiraz. Iran South Med J 2011; 14(2): 76-83.
  6. Brzuszkiewicz E, Thurmer A, Schuldes J, Leimbach A, Liesegang H, Meyer FD, et al. Genome sequence analyses of two isolates from the recent Escherichia coli outbreak in Germany reveal the emergence of a new pathotype: Entero-Aggregative-Haemorrhagic Escherichia coli (EAHEC). Arch Microbiol 2011; 193(12): 883-91.
  7. Kargar M, Homayoon M. Outbreaks of infection sources and antibiotic resistance in EHEC strains among children under 5 years old in Marvdasht. Med Sci J Islamic Azad Univ Tehran Med Branch 2010; 19(4): 268-73.
  8. Beddoe T, Paton AW, Le NJ, Rossjohn J, Paton JC. Structure, biological functions and applications of the AB5 toxins. Trends Biochem Sci 2010; 35(7): 411-8.
  9. Croxen MA, Finlay BB. Molecular mechanisms of Escherichia coli pathogenicity. Nat Rev Microbiol 2010; 8(1): 26-38.
  10. Shah Ili M, Kargar M, Rezaeian Jahrommi AA, Homayoon M, Kargar M, Ghorbani-Dalini S. Evaluation of virulance genes of shiga toxin producing Escherichia coli from juice purchase and vegetables in Shiraz. Journal of Microbial World 2010; 3 (1): 40-7.
  11. Karmali MA, Mascarenhas M, Shen S, Ziebell K, Johnson S, Reid-Smith R, et al. Association of genomic O island 122 of Escherichia coli EDL 933 with verocytotoxin-producing Escherichia coli seropathotypes that are linked to epidemic and/or serious disease. J Clin Microbiol 2003; 41(11): 4930-40.
  12. Smith HR, Scotland SM. ACP Broadsheet 135: January 1993. Isolation and identification methods for Escherichia coli O157 and other Vero cytotoxin producing strains. J Clin Pathol 1993; 46(1): 10-7.
  13. Gould LH, Bopp C, Strockbine N, Atkinson R, Baselski V, Body B, et al. Recommendations for diagnosis of shiga toxin--producing Escherichia coli infections by clinical laboratories. MMWR Recomm Rep 2009; 58(RR-12): 1-14.
  14. Viazis S, Diez-Gonzalez F. Enterohemorrhagic Escherichia coli: the twentieth century's emerging foodborne pathogen: a review. Advances in Agronomy 2011; 111: 1-50.
  15. Shams S, Haghi-Ashtiani MT, Nasrollahi L, Shahsiah R, Monajemzadeh M, Tahbaz-Lahafi B, et al. Frequency of shiga toxin-producing genes of Escherichia coli isolated from diarrheic stools of Iranian children by PCR. Iran J Pediatr 2013; 23(6): 637-42.
  16. Kargar M, Heidary S, Abbasian F, Shekarforoosh Sh. Survey of different enrichment methods, prevalence and antibiotic resistance of E. coli O157:H7 in raw milk of Jahrom cows. Iran J Infect Dis Trop Med 2006; 11(34): 7-11.
  17. Guion CE, Ochoa TJ, Walker CM, Barletta F, Cleary TG. Detection of diarrheagenic Escherichia coli by use of melting-curve analysis and real-time multiplex PCR. J Clin Microbiol 2008; 46(5): 1752-7.
  18. Vidal M, Kruger E, Duran C, Lagos R, Levine M, Prado V, et al. Single multiplex PCR assay to identify simultaneously the six categories of diarrheagenic Escherichia coli associated with enteric infections. J Clin Microbiol 2005; 43(10): 5362-5.
  19. Lopez-Saucedo C, Cerna JF, Villegas-Sepulveda N, Thompson R, Velazquez FR, Torres J, et al. Single multiplex polymerase chain reaction to detect diverse loci associated with diarrheagenic Escherichia coli. Emerg Infect Dis 2003; 9(1): 127-31.
  20. Wang L, Li Y, Mustaphai A. Rapid and simultaneous quantitation of Escherichia coli 0157:H7, Salmonella, and Shigella in ground beef by multiplex real-time PCR and immunomagnetic separation. J Food Prot 2007; 70(6): 1366-72.
  21. Silley P. Susceptibility testing methods, resistance and breakpoints: what do these terms really mean? Rev Sci Tech 2012; 31(1): 33-41.
  22. Performance standards for antimicrobial susceptibility testing; Twenty-first informational supplement. CLSI document M100-S21. Wayne, PA: Clinical and Laboratory Standards Institute; 2011.
  23. Keskimaki M, Eklund M, Pesonen H, Heiskanen T, Siitonen A. EPEC, EAEC and STEC in stool specimens: prevalence and molecular epidemiology of isolates. Diagn Microbiol Infect Dis 2001; 40(4): 151-6.
  24. Nessa K, Dilruba A, Islam J, Kabir L, Hossai MA. Usefulness of a multiplex PCR for detection of diarrheagenic escherichia coli in a diagnostic microbiology laboratory setting, Bangladesh. J Med Microbiol 2007; 1(2): 38-42.
  25. Alikhani MY, Mirsalehian A, Fatollahzadeh B, Pourshafie MR, Aslani MM. Prevalence of enteropathogenic and shiga toxin-producing Escherichia coli among children with and without diarrhoea in Iran. J Health Popul Nutr 2007; 25(1): 88-93.
  26. Jafari F, Garcia-Gil LJ, Salmanzadeh-Ahrabi S, Shokrzadeh L, Aslani MM, Pourhoseingholi MA, et al. Diagnosis and prevalence of enteropathogenic bacteria in children less than 5 years of age with acute diarrhea in Tehran children's hospitals. J Infect 2009; 58(1): 21-7.
  27. Jafari F, Shokrzadeh L, Hamidian M, Salmanzadeh-Ahrabi S, Zali MR. Acute diarrhea due to enteropathogenic bacteria in patients at hospitals in Tehran. Jpn J Infect Dis 2008; 61(4): 269-73.
  28. Byrne CM, Erol I, Call JE, Kaspar CW, Buege DR, Hiemke CJ, et al. Characterization of Escherichia coli O157:H7 from downer and healthy dairy cattle in the upper Midwest region of the United States. Appl Environ Microbiol 2003; 69(8): 4683-8.
  29. Alborzi A, Aelami MH, Astaneh B, Pourabbas B, Farshad S, Kalani M, et al. Is Escherichia coli O157:H7 a common pathogen in children with bloody diarrhea in Shiraz, Iran? Turk J Pediatr 2008; 50(4): 349-53.
  30. Nahaei MR, Akbari Dibavar M, Sadeghi J, Nikvash S. Frequency of enterohaemorrhagic Escherichia coli isolated from patients with acute diarrhea in Tabriz hospitals. Iranian J Med Microbiol 2007; 1(3): 39-46.
  31. Akbari A, Pourmand MR, Fard Sanei F, Mardani N, Soltan Dallal MM. Detection of E.coli strains containing shiga toxin (Stx1/2) gene in diarrheal specimens from children less than 5 years old by PCR technique and study of the patterns of antibiotic resistance. J Shaheed Sadoughi Univ Med Sci 2009, 17(4): 279-85.
Journal of Isfahan Medical School
Volume 32, Issue 273 - Serial Number 273
January and February 2014
Pages 67-78

Files

History

  • Receive Date: 28 August 2013
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics