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Updated: Oct 9, 2025

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Gut microbiota, innate immune pathways, and inflammatory control mechanisms in patients with major depressive
Javier R Caso1,2, Karina S MacDowell1,2, Ana González-Pinto3
1Department of Pharmacology & Toxicology, Faculty of Medicine, Universidad Complutense de Madrid (UCM), Madrid, Spain.
Abstract:
Although alterations in the gut microbiota have been linked to the pathophysiology of major depressive disorder (MDD), including through effects on the immune response, our understanding is deficient about the straight connection patterns among microbiota and MDD in patients. Male and female MDD patients were recruited: 46 patients with a current active MDD (a-MDD) and 22 in remission or with only mild symptoms (r-MDD). Forty-five healthy controls (HC) were also recruited. Psychopathological states were assessed, and fecal and blood samples were collected. Results indicated that the inducible nitric oxide synthase expression was higher in MDD patients compared with HC and the oxidative stress levels were greater in the a-MDD group. Furthermore, the lipopolysaccharide (an indirect marker of bacterial translocation) was higher in a-MDD patients compared with the other groups. Fecal samples did not cluster according to the presence or the absence of MDD. There were bacterial genera whose relative abundance was altered in MDD: Bilophila (2-fold) and Alistipes (1.5-fold) were higher, while Anaerostipes (1.5-fold) and Dialister (15-fold) were lower in MDD patients compared with HC. Patients with a-MDD presented higher relative abundance of Alistipes and Anaerostipes (1.5-fold) and a complete depletion of Dialister compared with HC. Patients with r-MDD presented higher abundance of Bilophila (2.5-fold) compared with HC. Thus, the abundance of bacterial genera and some immune pathways, both with potential implications in the pathophysiology of depression, appear to be altered in MDD, with the most noticeable changes occurring in patients with the worse clinical condition, the a-MDD group.
Insights
Major depressive disorder (MDD) is linked to gut bacteria changes, with specific genera like Dialister depleted in patients. These microbial shifts, particularly in active MDD, may influence depression pathophysiology and immune responses.
Area of Science:
- Microbiome research
- Neuroscience
- Immunology
Background:
- Alterations in gut microbiota are implicated in major depressive disorder (MDD) pathophysiology.
- The direct relationship between gut microbiota and MDD remains unclear, particularly concerning immune system involvement.
Purpose of the Study:
- To investigate the specific patterns of gut microbiota and their association with major depressive disorder (MDD).
- To explore the link between microbial alterations, immune markers, and the severity of depressive symptoms.
Main Methods:
- Recruited male and female patients with active MDD (a-MDD), in remission/mild MDD (r-MDD), and healthy controls (HC).
- Assessed psychopathological states and collected fecal and blood samples for analysis.
- Quantified inducible nitric oxide synthase, oxidative stress, lipopolysaccharide, and bacterial genus abundance.
Main Results:
- MDD patients showed higher inducible nitric oxide synthase expression and oxidative stress compared to HC.
- Lipopolysaccharide levels were elevated in a-MDD patients, suggesting bacterial translocation.
- Specific bacterial genera (Bilophila, Alistipes) were increased, while others (Anaerostipes, Dialister) were decreased in MDD patients, with Dialister notably depleted in a-MDD.
Conclusions:
- Gut microbiota composition and immune pathways are altered in MDD patients.
- The most significant microbial changes were observed in patients with active MDD, correlating with clinical severity.
- These findings suggest a role for gut microbial dysbiosis in the pathophysiology of depression.
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