Mètodes moleculars per l’estudi de comunitats microbianes

Abstract

This work presents a comprehensive review of key molecular techniques applied to the study

of microbial communities, particularly emphasizing experimental procedures, technical

particularities, specific advantages, and their suitability to different microbial contexts.

Metagenomics, based on total DNA analysis, enables the identification of microorganisms

present and characterizes the overall functional potential of complex or unknown communities,

providing high taxonomic and functional resolution. Metabarcoding, also DNA-based, employs

specific genetic markers (e.g., the 16S rRNA gene), allowing rapid and precise taxonomic

identification, ideal for highly diverse communities. Metatranscriptomics, centered on RNA

analysis, reveals actively expressed genes, providing valuable dynamic and functional

information for environmental or intestinal microbiome studies. Metaproteomics examines

proteins directly, informing about active metabolic functions, particularly effective when prior

genomic or metagenomic data is available. Metabolomics analyzes final microbial metabolites,

providing detailed physiological insight, especially suited to well-defined communities with

existing references. RT-qPCR specifically quantifies the expression of targeted genes with

high sensitivity, ideal for monitoring specific genes in targeted studies. Lastly, single-cell

genomics enables investigation of rare, uncultivable, or poorly understood microorganisms,

facilitating detailed genetic and functional characterization. This review emphasizes the

importance of selecting appropriate methodologies based on the specific characteristics of

each microbial community, highlighting the integration of these omics techniques as the

optimal strategy for a deep understanding of microbiomes

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