What is the role of fecal immunoassay testing in diagnosing gastrointestinal infections, such as Clostridioides (C. difficile) difficile?

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Last updated: August 29, 2025View editorial policy

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Role of Fecal Immunoassay Testing in Diagnosing C. difficile Infection

Fecal immunoassay testing should be used as part of a two-step algorithm for diagnosing Clostridioides difficile infection (CDI), with enzyme immunoassays (EIAs) for glutamate dehydrogenase (GDH) as an initial screening test followed by toxin A/B detection for confirmation. 1

Diagnostic Approach for C. difficile

When to Test

  • Test only unformed stool samples from symptomatic patients with ≥3 unformed stools in 24 hours 1
  • Consider testing in:
    • Patients >2 years of age with diarrhea following antimicrobial use
    • Patients with healthcare-associated diarrhea
    • Patients with persistent diarrhea without an identified cause 1

Specimen Collection

  • Fresh diarrheal stool sample (takes the shape of the container) is the preferred specimen 1
  • For patients with suspected severe CDI complicated by ileus who cannot produce stool specimens, perirectal swabs may be used (sensitivity 95.7%, specificity 100%) 1
  • A single stool specimen is sufficient; multiple specimens do not increase diagnostic yield 1

Testing Methods and Their Characteristics

Two-Step Algorithm

  1. First step: GDH EIA (high sensitivity screening test)
  2. Second step: Toxin A/B EIA (confirmation test)

Test Performance Characteristics

Test Sensitivity Specificity Advantages Disadvantages
GDH EIA High Moderate Excellent screening test Cannot differentiate toxigenic strains
Toxin A/B EIA 32-98% 84-100% Fast, inexpensive Variable sensitivity
NAATs 80-100% 87-99% High sensitivity, rapid results May detect colonization, not just infection
Toxigenic culture High High Gold standard Slow, labor-intensive

Test Interpretation

  • Negative first test: Report as negative for CDI 1
  • Positive first test + positive confirmatory test: Report as positive for CDI 1
  • Positive first test + negative confirmatory test: Cannot differentiate between infection and colonization 1

Common Pitfalls in CDI Diagnosis

  1. Testing formed stool: Only test unformed stool samples, as C. difficile can colonize without causing disease 1

  2. Over-reliance on single tests: No single test is perfect; laboratory results must be interpreted in the context of clinical presentation 1

  3. Confusing colonization with infection: A positive bacterial culture or NAAT without toxin detection may indicate colonization rather than infection 1

  4. Repeat testing during the same episode: A single stool specimen is sufficient; multiple specimens do not increase diagnostic yield 1

  5. Testing for cure: Avoid "test of cure" as patients may remain colonized after successful treatment 1

Clinical Context for Test Interpretation

The diagnosis of C. difficile infection requires both:

  1. Presence of compatible symptoms (typically diarrhea)
  2. Laboratory evidence of toxigenic C. difficile or its toxins in stool 1

Treatment Based on Diagnosis

Once CDI is confirmed through appropriate testing, treatment should be initiated:

  • First-line treatment: Fidaxomicin 200 mg twice daily for 10 days 1, 2
  • Alternative first-line: Vancomycin 125 mg four times daily for 10 days 1, 2
  • For non-severe CDI only (if first-line unavailable): Metronidazole 500 mg three times daily by mouth for 10-14 days 1, 2

Recent Developments

Recent studies emphasize the importance of proper diagnostic testing for CDI, with a shift away from metronidazole as first-line therapy 2, 3. The 2024 literature continues to support the two-step algorithm for diagnosis, highlighting that no single test should be used alone 4, 5, 3.

Fecal immunoassay testing remains a cornerstone in the diagnosis of C. difficile infection, providing rapid results when used appropriately as part of a comprehensive diagnostic approach.

References

Guideline

Diagnosing and Managing Clostridioides difficile Infection

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Research

When and What to Test for Diarrhea: Focus on Stool Testing.

The American journal of gastroenterology, 2024

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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