Search bioRxiv⌕ Search

Biology subjects

Branders, S.

Publications and source records attributed to Branders, S..

2 recordsLinked to original sources

Real-time Taxonomic Characterization of Long-read Mixed-species Sequencing Samples in Sorted Motif Distance Space: Voyager

Recent advances in long-read sequencing technology enable its use in potentially life-saving applications for rapid clinical diagnostics and epidemiological monitoring. To take advantage of these enabling characteristics, we present Voyager, a novel algorithm that complements real-time sequencing by rapidly and efficiently mapping long sequencing reads with insertion- and deletion errors to a large set of reference genomes. The concept of Sorted Motif Distance Space (SMDS), i.e., distances between exact matches of short motifs sorted by rank, represents sequences and sequence complementarity in a highly compressed form and is thus computationally efficient while enabling strain-level discrimination. In addition, Voyager applies a deconvolution algorithm rather than reducing taxonomic resolution if sequences of closely related organisms cannot be discerned by SMDS alone. Using relevant real-world data, we evaluated Voyager against the current best taxonomic classification methods (Kraken 2 and Centrifuge). Voyager was on average more than twice as fast as the current fastest method and obtained on average over 40% higher species level accuracy while maintaining lower memory usage than both other methods.

bioinformatics↗

Detection of pathogens and antimicrobial resistant genes from urine within 5 hours using Nanopore sequencing

PurposeUrinary Tract Infection (UTI) is a prevalent global health concern accounting for 1-3% of primary healthcare visits. The current methods for UTI diagnosis have a high turnaround time of 3-5 days for pathogen identification and susceptibility testing. This work is a proof-of-concept study aimed at determining the detection limit by establishing a culture and amplification-free DNA extraction methodology from spiked urine samples followed by real-time Nanopore sequencing and data analysis. MethodsThis study first establishes an optical density culture-based method for spiking healthy urine samples with the six most prevalent uropathogens. Pathogens were spiked at two clinically significant concentrations of 103 and 105 CFU/ml. Three commercial DNA extraction kits were investigated based on the quantity of isolated DNA, average processing time, elution volume and the average cost incurred per extraction. The outperforming kit was used for direct DNA extraction and subsequent sequencing on MinION and Flongle flowcells. ResultsThe Blood and Tissue kit outperformed the other kits. All pathogens were identified at a concentration of 105 CFU/ml within ten minutes, and the corresponding AMR genes were detected within three hours of the sequencing start. The overall turnaround time including the DNA extraction and sequencing steps was five hours. Moreover, we also demonstrate that the identification of some pathogens and antibiotic-resistance genes was possible at a spike concentration of 103 CFU/mL. ConclusionThis study shows great promise toward reducing the time required for making an informed antibiotic administration from approximately 48 hours to five hours thereby reducing the number of empirical doses and saving lives.

microbiology↗