Skip to main content Accessibility help
×
×
Home

Detection of Mixed Populations of Clostridium difficile from Symptomatic Patients Using Capillary-Based Polymerase Chain Reaction Ribotyping

  • Adam A. Behroozian (a1), Jeffrey P. Chludzinski (a1), Eugene S. Lo (a1), Sarah A. Ewing (a1), Sheila Waslawski (a1), Duane W. Newton (a2), Vincent B. Young (a1) (a3), David M. Aronoff (a1) (a3) and Seth T. Walk (a1) (a4)...

Abstract

Objective.

To investigate the simultaneous occurrence of more than 1 Clostridium difficile ribotype in patients' stool samples at the time of diagnostic testing.

Methods.

Stool samples submitted for diagnostic testing for the presence of toxigenic C. difficile were obtained for 102 unique patients. A total of 95 single colonies of C. difficile per stool sample were isolated on selective media, subcultured alongside negative (uninoculated) controls, and polymerase chain reaction (PCR) ribotyped using capillary gel electrophoresis.

Results.

Capillary-based PCR ribotyping was successful for 9,335 C. difficile isolates, yielding a median of 93 characterized isolates per stool sample (range, 69-95). More than 1 C. difficile ribotype was present in 16 of 102 (16%) C. difficile infection (CDI) cases; 2 of the 16 mixtures were composed of at least 3 ribotypes, while the remaining 14 were composed of at least 2.

Conclusions.

Deep sampling of patient stool samples coupled with capillary-based PCR ribotyping identified a high rate of mixed CDI cases compared with previous estimates. Studies seeking to quantify the clinical significance of particular C. difficile ribotypes should account for mixed cases of disease.

Copyright

Corresponding author

Montana State University, 109 Lewis Hall, Bozeman, MT 59717 (seth.walk@montana.edu)

References

Hide All
1.Kuijper, EJ, Oudbier, JH, Stuifbergen, WN, Jansz, A, Zanen, HC. Application of whole-cell DNA restriction endonuclease profiles to the epidemiology of Clostridium difficile-induced diarrhea. J Clin Microbiol 1987;25(4):751753.
2.Bidet, P, Barbut, F, Lalande, V, Burghoffer, B, Petit, JC. Development of a new PCR-ribotyping method for Clostridium difficile based on ribosomal RNA gene sequencing. FEMS Microbiol Lett 1999;175(2):261266.
3.Marsh, JW, O';Leary, MM, Shutt, KA, et al.Multilocus variable-number tandem-repeat analysis for investigation of Clostridium difficile transmission in hospitals. J Clin Microbiol 2006;44(7):25582566.
4.Eyre, DW, Walker, AS, Griffiths, D, et al.Clostridium difficile mixed infection and reinfection. J Clin Microbiol 2012;50(1):142144.
5.Hell, M, Permoser, M, Chmelizek, G, et al.Clostridium difficile infection: monoclonal or polyclonal genesis? Infection 2011; 39(5):461465.
6.van den Berg, RJ, Ameen, HA, Furusawa, T, Claas, EC, van der Vorm, ER, Kuijper, EJ. Coexistence of multiple PCR-ribotype strains of Clostridium difficile in faecal samples limits epidemiological studies. J Med Microbiol 2005;54(2):173179.
7.Wróblewski, D, Hannett, GE, Bopp, DJ, et al.Rapid molecular characterization of Clostridium difficile and assessment of populations of C. difficile in stool specimens. J Clin Microbiol 2009; 47(7):21422148.
8.Indra, A, Huhulescu, S, Schneeweis, M, et al.Characterization of Clostridium difficile isolates using capillary gel electrophoresis-based PCR ribotyping. J Med Microbiol 2008;57(11):13771382.
9.Walk, ST, Micie, D, Jain, R, et al.Clostridium difficile ribotype does not predict severe infection. Clin Infect Dis 2012;55(12):16611668.
10.McDonald, LC, Coignard, B, Dubberke, E, Song, X, Horan, T, Kutty, PK. Recommendations for surveillance of Clostridium difficile-associated disease. Infect Control Hosp Epidemiol 2007;28(2):140145.
11.Sorg, JA, Dineen, SS. Laboratory maintenance of Clostridium difficile. Curt Protoc Microbiol 2009;12:9A.1.19A.1.10.
12.Rinttila, T, Kassinen, A, Mahnen, E, Krogius, L, Palva, A. Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR. J Appi Microbiol 2004;97(6):11661177.
13.Persson, S, Torpdahl, M, Olsen, KE. 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(11):10571064.
14.Benjamini, Y, Yekutieli, D. The control of the false discovery rate in multiple testing under dependency. Ann Stat 2001;29(4):11651188.
15.Machin, D, Campbell, MJ, Fayers, PM, Pinol, APU. Sample Size Tables for Clinical Studies. 2nd ed. Oxford: Blackwell Science, 1997.
16.Balmer, O, Tanner, M. Prevalence and implications of multiple-strain infections. Lancet Infect Dis 2011;11(11):868878.
17.Bordello, SP, Honour, P. Concomitance of cytotoxigenic and non-cytotoxigenic Clostridium difficile in stool specimens. J Clin Microbiol 1983;18(4):10061007.
18.Sharp, J, Poxton, IR. An immunochemical method for fingerprinting Clostridium difficile. J Immunol Methods 1985;83(2):241248.
19.Barketi-Klai, A, Hoys, S, Lambert-Bordes, S, Collignon, A, Kansau, I. Role of fibronectin-binding protein A in Clostridium difficile intestinal colonization. J Med Microbiol 2011;60(8):11551161.
20.Merrigan, MM, Sambol, SP, Johnson, S, Gerding, DN. New approach to the management of Clostridium difficile infection: colonisation with non-toxigenic C. difficile during daily ampicillin or ceftriaxone administration. Int J Antimicrob Agents 2009; 33(suppl 1):S46S50.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Infection Control & Hospital Epidemiology
  • ISSN: 0899-823X
  • EISSN: 1559-6834
  • URL: /core/journals/infection-control-and-hospital-epidemiology
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Metrics

Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed