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Updated: May 2, 2026

Culturing and Maintaining Clostridium difficile in an Anaerobic Environment
Published on: September 15, 2013
Derivation and validation of guidelines for stool cultures for enteropathogenic bacteria other than Clostridium
T M Bauer1, A Lalvani, J Fehrenbach
1Department of Internal Medicine II, University Hospital Freiburg, Hugstetterstrasse 55, D-79106 Freiburg, Germany. bauert@med1.ukl.uni-freiburg.de
Context:
The yield of in-hospital stool cultures performed more than 72 hours after admission is low, and a commonly used policy dictates that laboratories reject these cultures to save costs. However, enteropathogenic bacteria other than Clostridium difficile (EPB) may cause nosocomial illness that would be missed by use of such a "3-day rule."
Objective:
To develop guidelines for hospital use of stool cultures that are sensitive to clinically relevant cases of sporadic and epidemic nosocomial diarrhea.
Design:
Five-part study that incorporated a derivation sample based on retrospective chart review and a prospective cohort study (including cost savings analysis), and a validation sample based on retrospective chart review.
Setting:
Four European academic health care centers.
Patients:
Derivation sample: 1735 adult inpatients from whom 3416 stool cultures were obtained during a 19-month period (1995-1997) and 68 adult inpatients for whom EPB were grown from stool cultures during a 10-year period (1988-1998); validation sample: 65 patients with sporadic isolation of EPB (1993-1998), 56 patients involved in 2 nosocomial Salmonella outbreaks (1992 and 1997), and 330 patients who had stool cultures performed (1998).
Main Outcome Measure:
Performance of derived criteria in detecting pathogenic bacteria and outbreaks and reducing total number of stool cultures performed.
Results:
Stool cultures grew EPB in 3.3% of samples obtained =72 hours after admission and 0.5% of samples obtained thereafter (P<.001). Isolation of EPB >72 hours after admission was not associated with clinical symptoms or signs but was associated with community-acquired diarrhea (24%), age 65 years or older with preexisting comorbid disease (25%), neutropenia (13%), HIV infection (10%), and nondiarrheal manifestations of enteric infections (16%). Twelve percent were asymptomatic carriers. These characteristics were used to create criteria for selecting patients for whom stool cultures would be indicated. These criteria were applied post hoc to a series of 1025 stool cultures; the number of stool cultures would have been reduced by 52% and no clinically significant cases would have been missed. Annual savings to a 355-bed institution would be approximately $7800 for reagent costs and 75 hours of technician time. In the validation samples, only 2 patients of 65 who had EPB would not have been identified, and neither required treatment. If the 3-day rule had been applied, 52 cases would not have been identified, 28 of which required antibiotic treatment.
Conclusion:
Our modified 3-day rule for use in selecting cases for stool culture is sensitive to sporadic and epidemic cases of nosocomial diarrhea in hospitalized adults.
Insights
A modified 3-day rule for hospital stool cultures effectively identifies enteropathogenic bacteria (EPB) and outbreaks, reducing unnecessary tests. This approach ensures sensitive detection of nosocomial diarrhea cases while improving cost-effectiveness.
Area of Science:
- Infectious Diseases
- Clinical Microbiology
- Hospital Epidemiology
Background:
- Hospital stool cultures performed >72 hours post-admission have low yield and are often rejected to cut costs.
- This
- 3-day rule
- policy may miss nosocomial infections caused by enteropathogenic bacteria other than Clostridium difficile (EPB).
Purpose of the Study:
- To develop guidelines for hospital stool culture use.
- Guidelines should be sensitive to clinically relevant sporadic and epidemic nosocomial diarrhea cases.
Main Methods:
- A five-part study involving derivation and validation samples from four European academic health care centers.
- Retrospective chart reviews and a prospective cohort study, including cost-saving analysis, were conducted.
- Criteria were derived from patient characteristics associated with EPB isolation >72 hours post-admission.
Main Results:
- EPB isolation was significantly lower >72 hours post-admission (0.5%) compared to =72 hours (3.3%).
- EPB isolation >72 hours was linked to community-acquired diarrhea, older age with comorbidities, neutropenia, HIV, and non-diarrheal enteric manifestations.
- Applying derived criteria reduced stool cultures by 52% with no missed clinically significant cases, yielding annual savings of ~$7800 and 75 technician hours.
Conclusions:
- A modified 3-day rule for stool culture selection is sensitive to sporadic and epidemic nosocomial diarrhea in hospitalized adults.
- This revised approach enhances diagnostic yield and cost-efficiency in hospital microbiology.
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