Critical Steps for Human Gut Exfoliome RNA Profiling Analysis Using Non-Invasive Stool Samples.
Introduction: Dietary exposure and drug treatments influence gut cellular pathways and hence growth and potentially even the gut-brain-microbiome axis. Since eukaryotic mRNA presents poly-A sequence that distinguishes them from the prokaryotes mRNA, we could analyze the gene expression of human gut...
| Publicado en: | Annals of Nutrition & Metabolism Vol. 78; no. 2; pp. 80 - 91 |
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| Autores principales: | , , , , |
| Formato: | pictorial research tables/charts tracings Journal Article |
| Publicado: |
Karger AG
2022
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=156200985&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 156200985 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 02506807 8BM jtl: Annals of Nutrition & Metabolism issn: 02506807 maglogo: N pubinfo: dt: 2022 vid: 78 iid: 2 pid: 2485 pub: Karger AG artinfo: ui: 156200985 156200985 156200985 10.1159/000520968 156200985 ppf: 80 ppct: 11 formats: tig: atl: Critical Steps for Human Gut Exfoliome RNA Profiling Analysis Using Non-Invasive Stool Samples. aug: au: Sánchez-Campillo, María Pastor-Fajardo, María Teresa Sabater-Molina, María López-Andreo, María José Larqué, Elvira affil: Department of Physiology, Faculty of Biology, Campus Mare Nostrum, University of Murcia, Murcia, Spain sug: subj: Gut Microbiota RNA, Messenger Sequence Analysis Feces Analysis Gene Expression Human Signal Transduction Microarray Analysis Biochips DNA Technology Utilization Epithelial Cells Bacterial Contamination ab: Introduction: Dietary exposure and drug treatments influence gut cellular pathways and hence growth and potentially even the gut-brain-microbiome axis. Since eukaryotic mRNA presents poly-A sequence that distinguishes them from the prokaryotes mRNA, we could analyze the gene expression of human gut cells using exfoliated gut cells available in stool samples. However, the impact of the critical steps of these non-invasive methods must be analyzed. Methods: We tested prokaryote contamination in all the steps of different procedures to analyze human exfoliome by microarrays and the influence of the fecal sampling collection process. Results: The least bacterial contamination was found using RNA amplified with oligo dT from the GeneChip 3′ IVT Pico Reagent Kit or using RNA purified by both Oligotex® + oligo dT. RNAlater® collection of feces affects the microarray results compared to directly frozen fecal samples, although both methods produce similar cDNA quality. Conclusion: This technique is a potential non-invasive diagnostic tool that can be applied to larger studies to quantify intestinal gene expression in humans with non-invasive samples, but samples should always be collected and analyzed under the same procedure. pubtype: Academic Journal doctype: pictorial research tables/charts tracings Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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