Modern approaches to gut microbiome investigation: Sequencing, culturomics, metabolomics, and beyond.

Abstract

The human gut microbiome is a complex and constantly evolving community of trillions of microorganisms that are crucial to various aspects of health and disease. It impacts digestion, metabolism, immune function, neurological processes, and vulnerability to illnesses. Recent technological advancements in biology and engineering have transformed microbiome research, allowing for more detailed analysis of microbial composition, functions, and interactions with the host. This review offers a thorough overview of both current and emerging methods for studying the gut microbiome, including sample collection techniques, culture-based approaches like culturomics and microfluidics, as well as culture-independent methods such as 16S rRNA sequencing, shotgun metagenomics, and the integration of multi-omics approaches like metabolomics, proteomics, and transcriptomics. It also discusses innovative tools including single-cell genomics, spatial transcriptomics, and microbiome-on-a-chip platforms, which hold promise for revealing host-microbe interactions at unprecedented levels of detail. The review underscores the importance of combining biological insights with engineering innovations particularly microfluidics and organ-on-a-chip models to recreate gut environments that mimic physiological conditions. Additionally, it explores the potential of artificial intelligence and machine learning in analyzing data and developing predictive models for personalized microbiome-based diagnostics and therapies. Acknowledging challenges such as microbial diversity, environmental sensitivity, and technical hurdles, this review aims to guide researchers in choosing optimal tools to study the gut microbiota, deepen mechanistic understanding, and translate findings into clinical applications that enhance human health.