Development of a Quantitative PCR Method for Detecting Enterococcus faecalis Cytolysin in Human Stool Samples

Area of Science:

• Gastroenterology and Hepatology
• Microbiology
• Molecular Diagnostics

Background:

• Alcohol-associated liver disease (ALD) is a significant global health concern.
• Alcohol-associated hepatitis (AH) presents high mortality and limited treatment options.
• Gut microbiota dysbiosis, specifically Enterococcus faecalis (E. faecalis) and its cytolysin, is implicated in ALD pathogenesis.

Purpose of the Study:

• To develop and standardize a real-time quantitative polymerase chain reaction (RT-qPCR) assay for detecting and quantifying cytolysin in fecal samples.
• To establish a diagnostic tool for identifying cytolysin-positive E. faecalis in patients with alcohol-associated hepatitis.
• To enable association analysis between cytolysin presence, disease severity, and mortality in AH.

Main Methods:

• Development of a standardized RT-qPCR assay.
• Detection and quantification of cytolysin in fecal samples.
• Correlation analysis between cytolysin detection and clinical outcomes (severity, mortality).

Main Results:

• A standardized RT-qPCR assay for cytolysin detection was successfully developed.
• The assay allows for the quantification of cytolysin in fecal samples from AH patients.
• The assay facilitates the analysis of the association between cytolysin-positive E. faecalis and disease severity/mortality.

Conclusions:

• The developed RT-qPCR assay provides a standardized method for identifying cytolysin-positive E. faecalis in alcohol-associated hepatitis.
• This diagnostic tool can aid in stratifying patients for clinical trials and understanding the role of E. faecalis cytolysin in AH.
• Further research can leverage this assay to explore targeted therapeutic strategies against E. faecalis in ALD.