Hostname: page-component-76fb5796d-qxdb6 Total loading time: 0 Render date: 2024-04-25T10:35:28.132Z Has data issue: false hasContentIssue false
  • English
  • Français

Immunological Stability of Clostridium difficile Toxins in Clinical Specimens

Published online by Cambridge University Press:  19 February 2018

Donna M. Schora ,
Lance R. Peterson  and
Elena A. Usacheva
Donna M. Schora
Affiliation:
Infectious Disease Research, NorthShore University HealthSystem, Evanston, Illinois
Lance R. Peterson
Affiliation:
Infectious Disease Research, NorthShore University HealthSystem, Evanston, Illinois Pritzker School of Medicine, University of Chicago, Chicago, Illinois
Elena A. Usacheva*
Affiliation:
Infectious Disease Research, NorthShore University HealthSystem, Evanston, Illinois Pritzker School of Medicine, University of Chicago, Chicago, Illinois
*
Address correspondence to Elena A. Usacheva, PhD, NorthShore University HealthSystem, 2650 Ridge Ave, Walgreen SB #525, Evanston, IL 60201, (EUsacheva@northshore.org).
Get access Rights & Permissions [Opens in a new window]

Abstract

OBJECTIVE

The impact of storage on stability and detection of Clostridium difficile toxins in feces is poorly understood. The objective of this study was to investigate the immunological stability of C. difficile toxins in clinical stool specimens under different storage conditions by evaluating this stability using toxin detection by enzyme immunoassay (EIA).

METHODS

Stool specimens positive for C. difficile infection (CDI) by quantitative polymerase chain reaction (qPCR) were used for EIA testing with the C. difficile Tox A/B II kit. The EIA-positive specimens were stored aerobically under refrigerated (4–10°C) and frozen (−30°C and −80°C) conditions. Measurement of toxin quantity was conducting using optical density (OD) on days 0, 14, 30, 60, 90, and 120 of storage.

RESULTS

Clostridium difficile toxins demonstrated good detection in undiluted stool specimens by EIA up to 120 days of storage. Good detection of the toxins was observed in diluted samples at refrigerated and −80°C temperatures. Dilution detrimentally affected toxin detection at −30°C.

CONCLUSION

Storage of undiluted clinical stool specimens at refrigerated, −30°C, and −80°C temperatures for up to 120 days has no discernible effect on the immunological stability of C. difficile cytotoxins. However, storage at −30°C has a detrimental effect on C. difficile toxin stability in diluted specimens.

Infect Control Hosp Epidemiol 2018;39:434–438

Type
Original Articles
Information
Infection Control & Hospital Epidemiology , Volume 39 , Issue 4 , April 2018 , pp. 434 - 438
Copyright
© 2018 by The Society for Healthcare Epidemiology of America. All rights reserved 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

PREVIOUS PRESENTATION. These results were presented in part at the Tenth International Society for Applied Biological Sciences (ISABS) Conference on June 19–24, 2017, in Dubrovnik, Republic of Croatia.

References

REFERENCES

1. Burke, KE, Lamont, JT. Clostridium difficile infection: a worldwide disease. Gut Liver 2014;8:1–6.CrossRefGoogle ScholarPubMed
2. Carter, GP, Chakravorty, A, Pham Nguyen, TA, et al. Defining the roles of TcdA and TcdB in localized gastrointestinal disease, systemic organ damage, and the host response during Clostridium difficile infections. MBio 2015;6:e00551.Google Scholar
3. Usacheva, EA, Jin, JP, Peterson, LR. Host response to Clostridium difficile infection: diagnostics and detection. J Glob Antimicrob Resist 2016;7:93–101.CrossRefGoogle ScholarPubMed
4. Burnham, CA, Carroll, KC. Diagnosis of Clostridium difficile infection: an ongoing conundrum for clinicians and for clinical laboratories. Clin Microbiol Rev 2013;26:604–630.Google Scholar
5. Davis, MY, Zhang, H, Brannan, LE, Carman, RJ, Boone, JH. Rapid change of fecal microbiome and disappearance of Clostridium difficile in a colonized infant after transition from breast milk to cow milk. Microbiome 2016;4:53.CrossRefGoogle Scholar
6. Butler, M, Olson, A, Drekonja, D, et al. Early diagnosis, prevention, and treatment of Clostridium difficile: update. Comparative effectiveness reviews, no. 172. Agency for Healthcare Research and Quality website. https://www.ncbi.nlm.nih.gov/books/NBK361173/. Published March 2016. Accessed January 22, 2018.Google Scholar
7. Healthcare-associated infections. Frequently asked questions about Clostridium difficile for healthcare providers. Centers for Disease Control and Prevention website. http://www.cdc.gov/HAI/organisms/cdiff/Cdiff_faqs_HCP.html. Published 2010; updated 2012. Accessed January 22, 2018.Google Scholar
8. Stevens, D, Bryant, A, Carroll, K. Clostriduim. Vol 11 Washington, DC: ASM Press; 2015.Google Scholar
9. Freeman, J, Wilcox, MH. The effects of storage conditions on viability of Clostridium difficile vegetative cells and spores and toxin activity in human faeces. J Clin Pathol 2003;56:126–128.Google Scholar
10. Weese, JS, Staempfli, HR, Prescott, JF. Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation. J Vet Diagn Invest 2000;12:332–336.CrossRefGoogle ScholarPubMed
11. Bowman, RA, Riley, TV. Isolation of Clostridium difficile from stored specimens and comparative susceptibility of various tissue culture cell lines to cytotoxin. FEMS Lett 1986;34:31–35.Google Scholar
12. Becker, SL, Chatigre, JK, Coulibaly, JT, et al. Molecular and culture-based diagnosis of Clostridium difficile isolates from Cote d’Ivoire after prolonged storage at disrupted cold chain conditions. Trans R Soc Trop Med Hyg 2015;109:660–668.Google Scholar
13. Modi, C, DePasquale, JR, Nguyen, NQ, Malinowski, JE, Perez, G. Does the handling time of unrefrigerated human fecal specimens impact the detection of Clostridium difficile toxins in a hospital setting? Indian J Gastroenterol 2010;29:157–161.Google Scholar
14. Corthier, G, Muller, MC, Elmer, GW, Lucas, F, Dubos-Ramare, F. Interrelationships between digestive proteolytic activities and production and quantitation of toxins in pseudomembranous colitis induced by Clostridium difficile in gnotobiotic mice. Infect Immun 1989;57:3922–3927.CrossRefGoogle ScholarPubMed
15. Lei, XH, Bochner, BR. Using phenotype microarrays to determine culture conditions that induce or repress toxin production by Clostridium difficile and other microorganisms. PLoS One 2013;8:e56545.Google Scholar
16. Valdes, L, Gueimonde, M, Ruas-Madiedo, P. Monitoring in real time the cytotoxic effect of Clostridium difficile upon the intestinal epithelial cell line HT29. J Microbiol Methods 2015;119:66–73.Google Scholar