ON-Time is described, a method for rapid and accurate metagenomic sequencing and taxonomic analysis of microbiomes in individual patient fecal samples, called ON-Time, which uses a simplified and rapid wet-lab workflow coupled with point-and-click data analysis.
Abstract
Clinical application of microbiome-guided therapies in the intensive care unit (ICU) requires a method to rapidly analyze patient microbiomes to guide urgent treatment decisions. Conventional microbiome sequencing and analysis methods require long turnaround times, methodological complexity, and high costs that are barriers to clinical application. Here, we describe a method for rapid (<5 h turnaround from sample to results) and accurate metagenomic sequencing and taxonomic analysis of microbiomes in individual patient fecal samples, called ON-Time. ON-Time uses a simplified and rapid wet-lab workflow coupled with point-and-click data analysis. Accuracy and precision of ON-Time data were validated using defined mock microbial communities and head-to-head comparison with conventional shotgun metagenomics of ICU patient samples. Key limitations include stochastic identification of functional genes such as antimicrobial resistance and virulence factors. Taken together, ON-Time offers a rapid, accurate, and cost-effective method to analyze individual patient samples for clinically actionable microbiome features to guide personalized therapeutics.
Advances in sequencing technologies have enabled increasingly detailed characterisation of the human microbiome in clinical studies, but interpretation of microbiome modulation which has relevance to health and disease characterisation or safety assessments remains methodologically challenging. Taxonomic profiles gener...
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Background: Precision diagnostics are more and more important in intensive care units (ICUs), where rapid identification of infectious agents and antimicrobial resistance determinants is crucial for timely and appropriate treatment. Conventional microbiological methods are frequently limited by long turnaround times an...
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Four types of bead-based mock communities comprising 26 species commonly found in the gastrointestinal tract, respiratory tract, skin, and genital tract plus cerebrospinal fluid are developed and are essential QC materials for clinical metagenomic NGS analysis.
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