Análisis del microbioma intestinal en pacientes pediátricos mediante secuenciación de nueva generación (NGS) y su posible relación con la carga de genes de resistencia a carbapenémicos

Abstract

The human gut microbiome plays a crucial role in pediatric development and immune modulation. Factors such as birth mode, feeding and antibiotic exposure have significant impact on the development and establishment of the microbiome. This study aimed to analyze the diversity of the intestinal microbiome in pediatric patients and evaluate its potential association with the load of carbapenem resistance genes. Rectal swab samples (n=46) from patients under 18 years of age who received healthcare at Hospital Universitario La Paz (HULP, Madrid, Spain) were analyzed. The results of quantitative PCRs were used to determine the relative abundance of resistance genes (blaKPC, blaNDM, blaOXA-48 and blaVIM) normalized to 16S rRNA gene levels. Samples were categorized into high, medium and negative intestinal relative load groups based on qPCR results. Following sample selection and classification, DNA was extracted and a full-length 16S rRNA gene was amplified by end point PCR to enable subsequent library preparation for sequencing using Oxford Nanopore Technology.