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Updated: Jan 16, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Gut Microbiota as a Key Player in Health and Disease with a Focus on Long-Term Alterations in Post-COVID
Ana Bačić1, Tijana Gmizić2, Tea Takić1
1Innovation Centre of Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, Serbia.
Introduction:
Gut microbiota plays a critical role in health and may contribute to post-COVID manifestations, as alterations in microbial communities have been observed in affected individuals. This study aimed to identify microbial and biochemical signatures in post-COVID individuals after an extended period following acute infection.
Methods:
A total of 90 subjects were recruited, divided into three equal groups: post-COVID, fully recovered individuals, and healthy controls. The post-COVID group was further stratified by symptom duration and severity. Fecal microbiota was analyzed using 16S rRNA gene sequencing (V3-V4 region), with differential abundance assessed through LEfSE. Biochemical parameters were measured using the Mindray BS800 system. Spearman's correlation was applied to explore associations between taxonomic groups and biochemical markers.
Results:
Several microbial taxa exhibited positive correlations with high-density lipoprotein (HDL) levels and negative associations with non-HDL lipid parameters and CRP, highlighting their potential role in health. Individuals with a history of COVID-19 showed a significantly lower relative abundance of Phascolarctobacterium, Slackia, Enterorhabdus, and Senegalimassilia (p < 0.05, LDA >2) compared to healthy controls. In participants with post-COVID symptoms, Eggerthella was significantly enriched, with Holdemania, Gemella, and two uncultured Lachnospiraceae showing a trend toward higher abundance. Severe post-COVID cases showed higher Parasutterella abundance, while long-lasting symptoms (>1 year) were associated with Dialister enrichment and Phascolarctobacterium depletion compared to controls and fully recovered individuals.
Conclusion:
Gut microbiota alterations were observed in post-COVID individuals even after a prolonged period following acute infection, suggesting a potential link between microbial imbalance and lingering symptoms, though causality cannot be confirmed.
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