Archives of Pediatric Infectious Diseases

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Comparison of the Detection Limits of the Culture and PCR Methods for the Detection of Clostridium difficile, Clostridium perfringens, Campylobacter jejuni, and Yersinia enterocolitica in Human Stool

Leila Ganji 1 , Masoumeh Azimirad 2 , Nastaran Farzi 2 , 3 , Masoud Alebouyeh 2 , 3 , * , Mohammad Hassan Shirazi 1 , * , Seyed Saeed Eshraghi 1 , Abbas Mirshafiey 1 , Naser Ebrahimi Daryani 1 and Mohammad Reza Zali 2 , 3
Authors Information
1 Department of Pathobiology, School of Public Health, University of Medical Sciences, Tehran, Iran
2 Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
3 Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Corresponding Authors:
Article information
  • Archives of Pediatric Infectious Diseases: January 01, 2017, 5 (1); e38888
  • Published Online: September 4, 2016
  • Article Type: Research Article
  • Received: May 1, 2016
  • Revised: July 20, 2016
  • Accepted: August 2, 2016
  • DOI: 10.5812/pedinfect.38888

To Cite: Ganji L, Azimirad M, Farzi N, Alebouyeh M, Shirazi M H, et al. Comparison of the Detection Limits of the Culture and PCR Methods for the Detection of Clostridium difficile, Clostridium perfringens, Campylobacter jejuni, and Yersinia enterocolitica in Human Stool, Arch Pediatr Infect Dis. 2017 ; 5(1):e38888. doi: 10.5812/pedinfect.38888.

Copyright © 2016, Pediartric Infections Research Center. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License ( which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
1. Background
2. Objectives
3. Methods
4. Results
5. Discussion
  • 1. Gerritsen J, Smidt H, Rijkers GT, de Vos WM. Intestinal microbiota in human health and disease: the impact of probiotics. Genes Nutr. 2011; 6(3): 209-40[DOI][PubMed]
  • 2. Guarner F, Malagelada JR. Gut flora in health and disease. Lancet. 2003; 361(9356): 512-9
  • 3. Hofreuter D, Tsai J, Watson RO, Novik V, Altman B, Benitez M, et al. Unique features of a highly pathogenic Campylobacter jejuni strain. Infect Immun. 2006; 74(8): 4694-707[DOI][PubMed]
  • 4. Allos BM. Campylobacter jejuni Infections: update on emerging issues and trends. Clin Infect Dis. 2001; 32(8): 1201-6[DOI][PubMed]
  • 5. Allos BM, Moore MR, Griffin PM, Tauxe RV. Surveillance for sporadic foodborne disease in the 21st century: the FoodNet perspective. Clin Infect Dis. 2004; 38 Suppl 3: 115-20[DOI][PubMed]
  • 6. Ashbolt NJ. Microbial contamination of drinking water and disease outcomes in developing regions. Toxicology. 2004; 198(1-3): 229-38[DOI][PubMed]
  • 7. Bartlett JG. Narrative review: the new epidemic of Clostridium difficile-associated enteric disease. Ann Intern Med. 2006; 145(10): 758-64[PubMed]
  • 8. Leggett HC, Cornwallis CK, West SA. Mechanisms of pathogenesis, infective dose and virulence in human parasites. PLoS Pathog. 2012; 8(2): 1002512[DOI][PubMed]
  • 9. Coker AO, Isokpehi RD, Thomas BN, Amisu KO, Obi CL. Human campylobacteriosis in developing countries. Emerg Infect Dis. 2002; 8(3): 237-44[DOI][PubMed]
  • 10. Olsen JE, Aabo S, Hill W, Notermans S, Wernars K, Granum PE, et al. Probes and polymerase chain reaction for detection of food-borne bacterial pathogens. Int J Food Microbiol. 1995; 28(1): 1-78[PubMed]
  • 11. Sloan LM, Duresko BJ, Gustafson DR, Rosenblatt JE. Comparison of real-time PCR for detection of the tcdC gene with four toxin immunoassays and culture in diagnosis of Clostridium difficile infection. J Clin Microbiol. 2008; 46(6): 1996-2001[DOI][PubMed]
  • 12. Oliver JD. The viable but nonculturable state in bacteria. J Microbiol. 2005; 43 Spec No: 93-100[PubMed]
  • 13. Talibart R, Denis M, Castillo A, Cappelier JM, Ermel G. Survival and recovery of viable but noncultivable forms of Campylobacter in aqueous microcosm. Int J Food Microbiol. 2000; 55(1-3): 263-7[PubMed]
  • 14. Iijima Y, Asako NT, Aihara M, Hayashi K. Improvement in the detection rate of diarrhoeagenic bacteria in human stool specimens by a rapid real-time PCR assay. J Med Microbiol. 2004; 53: 617-22[DOI][PubMed]
  • 15. Alvarez J, Sota M, Vivanco AB, Perales I, Cisterna R, Rementeria A, et al. Development of a multiplex PCR technique for detection and epidemiological typing of salmonella in human clinical samples. J Clin Microbiol. 2004; 42(4): 1734-8[PubMed]
  • 16. Collins E, Glennon M, Hanley S, Murray AM, Cormican M, Smith T, et al. Evaluation of a PCR/DNA probe colorimetric membrane assay for identification of Campylobacter spp. in human stool specimens. J Clin Microbiol. 2001; 39(11): 4163-5[DOI][PubMed]
  • 17. Wang G, Clark CG, Taylor TM, Pucknell C, Barton C, Price L, et al. Colony multiplex PCR assay for identification and differentiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus subsp. fetus. J Clin Microbiol. 2002; 40(12): 4744-7[PubMed]
  • 18. Yang C, Jiang Y, Huang K, Zhu C, Yin Y. Application of real-time PCR for quantitative detection of Campylobacter jejuni in poultry, milk and environmental water. FEMS Immunol Med Microbiol. 2003; 38(3): 265-71[PubMed]
  • 19. Clermont O, Bonacorsi S, Bingen E. Rapid and simple determination of the Escherichia coli phylogenetic group. Appl Environ Microbiol. 2000; 66(10): 4555-8[PubMed]
  • 20. Stamper PD, Alcabasa R, Aird D, Babiker W, Wehrlin J, Ikpeama I, et al. Comparison of a commercial real-time PCR assay for tcdB detection to a cell culture cytotoxicity assay and toxigenic culture for direct detection of toxin-producing Clostridium difficile in clinical samples. J Clin Microbiol. 2009; 47(2): 373-8[DOI][PubMed]
  • 21. Aichinger E, Schleck CD, Harmsen WS, Nyre LM, Patel R. Nonutility of repeat laboratory testing for detection of Clostridium difficile by use of PCR or enzyme immunoassay. J Clin Microbiol. 2008; 46(11): 3795-7[DOI][PubMed]
  • 22. Jones DM, Sutcliffe EM, Curry A. Recovery of viable but non-culturable Campylobacter jejuni. J Gen Microbiol. 1991; 137(10): 2477-82[DOI][PubMed]
  • 23. Burd EM. Validation of laboratory-developed molecular assays for infectious diseases. Clin Microbiol Rev. 2010; 23(3): 550-76[DOI][PubMed]
  • 24. Onori M, Coltella L, Mancinelli L, Argentieri M, Menichella D, Villani A, et al. Evaluation of a multiplex PCR assay for simultaneous detection of bacterial and viral enteropathogens in stool samples of paediatric patients. Diagn Microbiol Infect Dis. 2014; 79(2): 149-54[DOI][PubMed]
  • 25. Stenkova AM, Isaeva MP, Rasskazov VA. [Development of a multiplex PCR for detection of the Yersinia genus with identification of pathogenic species (Y. pestis, Y. pseudotuberculosis, Y. enterocolitica)]. Mol Gen Mikrobiol Virusol. 2008; (3): 18-23[PubMed]
  • 26. Spigaglia P, Mastrantonio P. Molecular analysis of the pathogenicity locus and polymorphism in the putative negative regulator of toxin production (TcdC) among Clostridium difficile clinical isolates. J Clin Microbiol. 2002; 40(9): 3470-5[PubMed]
  • 27. Wu J, Zhang W, Xie B, Wu M, Tong X, Kalpoe J, et al. Detection and toxin typing of Clostridium perfringens in formalin-fixed, paraffin-embedded tissue samples by PCR. J Clin Microbiol. 2009; 47(3): 807-10[DOI][PubMed]
  • 28. Jalali M, Khorvash F, Warriner K, Weese JS. Clostridium difficile infection in an Iranian hospital. BMC Res Notes. 2012; 5: 159[DOI][PubMed]
  • 29. Dallal MMS, Doyle MP, Rezadehbashi M, Dabiri H, Sanaei M, Modarresi S, et al. Prevalence and antimicrobial resistance profiles of Salmonella serotypes, Campylobacter and Yersinia spp. isolated from retail chicken and beef, Tehran, Iran. Food Control. 2010; 21(4): 388-92
  • 30. Feizabadi MM, Dolatabadi S, Zali MR. Isolation and drug-resistant patterns of Campylobacter strains cultured from diarrheic children in Tehran. Jpn J Infect Dis. 2007; 60(4): 217-9[PubMed]
  • 31. Ricciardi R, Rothenberger DA, Madoff RD, Baxter NN. Increasing prevalence and severity of Clostridium difficile colitis in hospitalized patients in the United States. Arch Surg. 2007; 142(7): 624-31[DOI][PubMed]
  • 32. Singh H, Rathore RS, Singh S, Cheema PS. Comparative analysis of cultural isolation and PCR based assay for detection of campylobacter jejuni in food and faecal samples. Braz J Microbiol. 2011; 42(1): 181-6[DOI][PubMed]
  • 33. Persson S, Olsen KE. Multiplex PCR for identification of Campylobacter coli and Campylobacter jejuni from pure cultures and directly on stool samples. J Med Microbiol. 2005; 54: 1043-7[DOI][PubMed]
  • 34. Leblanc-Maridor M, Beaudeau F, Seegers H, Denis M, Belloc C. Rapid identification and quantification of Campylobacter coli and Campylobacter jejuni by real-time PCR in pure cultures and in complex samples. BMC Microbiol. 2011; 11: 113[DOI][PubMed]
  • 35. Potturi-Venkata LP, Backert S, Lastovica AJ, Vieira SL, Norton RA, Miller RS, et al. Evaluation of different plate media for direct cultivation of Campylobacter species from live broilers. Poult Sci. 2007; 86(7): 1304-11[PubMed]
  • 36. Banting GS, Braithwaite S, Scott C, Kim J, Jeon B, Ashbolt N, et al. Evaluation of Various Campylobacter-Specific Quantitative PCR (qPCR) Assays for Detection and Enumeration of Campylobacteraceae in Irrigation Water and Wastewater via a Miniaturized Most-Probable-Number-qPCR Assay. Appl Environ Microbiol. 2016; 82(15): 4743-56[DOI][PubMed]
  • 37. Orlofsky E, Benami M, Gross A, Dutt M, Gillor O. Rapid MPN-Qpcr screening for pathogens in air, soil, water, and agricultural produce. Water Air Soil Pollut. 2015; 226(9): 1-10
  • 38. Wiemer D, Loderstaedt U, von Wulffen H, Priesnitz S, Fischer M, Tannich E, et al. Real-time multiplex PCR for simultaneous detection of Campylobacter jejuni, Salmonella, Shigella and Yersinia species in fecal samples. Int J Med Microbiol. 2011; 301(7): 577-84[DOI][PubMed]
  • 39. Zheng HX, Zhang MJ, Sun Y, Jiang B. [Detection of Yersinia enterocolitica in diarrhea stool by real-time PCR]. Zhonghua Yi Xue Za Zhi. 2006; 86(32): 2281-4[PubMed]
  • 40. Boyapalle S, Wesley IV, Hurd HS, Reddy PG. Comparison of culture, multiplex, and 5' nuclease polymerase chain reaction assays for the rapid detection of Yersinia enterocolitica in swine and pork products. J Food Prot. 2001; 64(9): 1352-61[PubMed]
  • 41. Belanger SD, Boissinot M, Clairoux N, Picard FJ, Bergeron MG. Rapid detection of Clostridium difficile in feces by real-time PCR. J Clin Microbiol. 2003; 41(2): 730-4[PubMed]
  • 42. Wise MG, Siragusa GR. Quantitative detection of Clostridium perfringens in the broiler fowl gastrointestinal tract by real-time PCR. Appl Environ Microbiol. 2005; 71(7): 3911-6[DOI][PubMed]
  • 43. Luna RA, Boyanton BL, Mehta S, Courtney EM, Webb CR, Revell PA, et al. Rapid stool-based diagnosis of Clostridium difficile infection by real-time PCR in a children's hospital. J Clin Microbiol. 2011; 49(3): 851-7[DOI][PubMed]
  • 44. Wannet WJ, Reessink M, Brunings HA, Maas HM. Detection of pathogenic Yersinia enterocolitica by a rapid and sensitive duplex PCR assay. J Clin Microbiol. 2001; 39(12): 4483-6[DOI][PubMed]
  • 45. Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson MA, Roy SL, et al. Foodborne illness acquired in the United States--major pathogens. Emerg Infect Dis. 2011; 17(1): 7-15[DOI][PubMed]
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