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Fecal micro RNA Isolation
Published on: October 28, 2020
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Critical Steps for Human Gut Exfoliome RNA Profiling Analysis Using Non-Invasive Stool Samples.
María Sánchez-Campillo1,2, María Teresa Pastor-Fajardo2, María Sabater-Molina2
1Department of Physiology, Faculty of Biology, Campus Mare Nostrum, University of Murcia, Murcia, Spain.
Annals of Nutrition & Metabolism
|November 17, 2021
Summary
Analyzing gene expression in human gut cells is possible using non-invasive stool samples. Minimizing bacterial contamination is key for accurate results from exfoliome analysis.
Area of Science:
- Gut microbiome research
- Molecular diagnostics
- Human gene expression analysis
Background:
- Diet and drugs impact gut pathways and the gut-brain-microbiome axis.
- Eukaryotic mRNA's poly-A sequence allows analysis of human gut cells from stool.
- Non-invasive methods for analyzing exfoliated gut cells require critical step evaluation.
Purpose of the Study:
- To assess prokaryotic contamination in human exfoliome analysis.
- To evaluate the influence of fecal sampling collection on microarray results.
- To optimize non-invasive methods for quantifying intestinal gene expression.
Main Methods:
- Microarray analysis of human exfoliome.
- Testing prokaryote contamination across different procedures.
- Evaluating the impact of fecal sampling collection methods.
Main Results:
- RNA amplification with oligo dT or purification with Oligotex® + oligo dT yielded minimal bacterial contamination.
- RNAlater® collection of feces significantly affected microarray results compared to direct freezing.
- Both collection methods produced comparable cDNA quality.
Conclusions:
- Human exfoliome analysis offers a potential non-invasive tool for intestinal gene expression studies.
- Standardized sample collection and analysis procedures are crucial for reliable results.
- This method can be applied to larger human studies for diagnostic purposes.
