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A Protein Microarray Assay for Serological Determination of Antigen-specific Antibody Responses Following Clostridium difficile Infection
Published on: June 15, 2018
Cytoskeletal Tropomyosin as a Biomarker in Clostridium difficile Infection
Elena A Usacheva1,2, Lance R Peterson1,2, Katherine Mendoza1
1Infectious Disease Research, NorthShore University HealthSystem, 2650 Ridge Ave., Evanston, IL 60201, USA.
Background:
Current diagnostics of Clostridium difficile infection (CDI) heavily relies on detection of the disease-causing organism. The objective of this study was to investigate a cytoskeletal protein, tropomyosin (Tpm), as a CDI biomarker.
Methods:
Fecal Tpm was tested by monoclonal antibodies (mAbs) in a 12-month prospective study. Remnant diarrheal clinical specimens and relevant clinical data were collected. The CDI positive (CDI+, n = 230) and CDI negative (CDI-, n = 228) groups were composed of samples testing positive or negative by polymerase chain reaction (PCR) (Xpert® C. difficile/Epi, Cepheid), respectively. The other enteric pathogen (OEP) group (n = 52) was composed of specimens tested for the presence of other enteric pathogens or parasites by routine testing methods. Extracted fecal Tpm was detected by Western blot and the results were correlated with CDI based on clinical and microbiology laboratory data.
Results:
A total of 510 stool specimens were tested. Tpm is not stable in stool, suggesting the utility of fresh specimens. In the CDI+ group, specificity and sensitivity of Tpm detection in correlation with a CDI were 93.2% and 53.7%, respectively, when only "true CDI" and "not CDI" were analyzed (110 samples). For CDI+ samples, 23% did not satisfy CDI clinical signs. Tpm positives in the CDI- group (8.3%) had inflammatory bowel diseases.
Conclusion:
Tpm has a potential role as a CDI biomarker in combination with C. difficile PCR and an appropriate clinical evaluation. However, non-muscle Tpm, as a biomarker for CDI, suffers from a low sensitivity in our study. Therefore further investigation using larger cohorts is needed.
Insights
Tropomyosin (Tpm) shows potential as a biomarker for Clostridium difficile infection (CDI), but its low sensitivity requires further study. Combining Tpm detection with PCR and clinical evaluation may improve CDI diagnostics.
Area of Science:
- Microbiology
- Clinical Diagnostics
- Biomarker Discovery
Background:
- Current Clostridium difficile infection (CDI) diagnostics primarily detect the organism.
- Investigating tropomyosin (Tpm), a cytoskeletal protein, as a novel CDI biomarker.
Purpose of the Study:
- To evaluate the diagnostic utility of fecal Tpm for Clostridium difficile infection (CDI).
- To assess the sensitivity and specificity of Tpm detection in stool specimens.
Main Methods:
- A prospective study analyzed 510 fecal specimens using monoclonal antibodies against Tpm.
- Samples were categorized as CDI positive (PCR-based) or negative, with an additional group for other enteric pathogens.
- Fecal Tpm levels were measured by Western blot and correlated with clinical and microbiological data.
Main Results:
- Tropomyosin (Tpm) stability in stool is limited, highlighting the need for fresh specimens.
- Tpm detection showed 93.2% specificity and 53.7% sensitivity for CDI.
- A subset of CDI positive samples (23%) lacked typical clinical signs, and Tpm positives in the CDI negative group (8.3%) had inflammatory bowel diseases.
Conclusions:
- Tropomyosin (Tpm) may serve as a supplementary biomarker for CDI when used with PCR and clinical assessment.
- Non-muscle Tpm demonstrated low sensitivity as a CDI biomarker in this study.
- Larger cohort studies are necessary to further validate Tpm's role in CDI diagnostics.
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