Abstract

BackgroundSequencing highly-variable 16S regions is a common and often effective approach to the study of microbial communities, and next-generation sequencing (NGS) technologies provide abundant quantities of data for analysis. However, the speed of existing analysis pipelines may limit our ability to work with these quantities of data. Furthermore, the limited coverage of existing 16S databases may hamper our ability to characterise these communities, particularly in the context of complex or poorly studied environments.ResultsIn this article we present the SigClust algorithm, a novel clustering method involving the transformation of sequence reads into binary signatures. When compared to other published methods, SigClust yields superior cluster coherence and separation of metagenomic read data, while operating within substantially reduced timeframes. We demonstrate its utility on published Illumina datasets and on a large collection of labelled wound reads sourced from patients in a wound clinic. The temporal analysis is based on tracking the dominant clusters of wound samples over time. The analysis can identify markers of both healing and non-healing wounds in response to treatment. Prominent clusters are found, corresponding to bacterial species known to be associated with unfavourable healing outcomes, including a number of strains of Staphylococcus aureus.ConclusionsSigClust identifies clusters rapidly and supports an improved understanding of the wound microbiome without reliance on a reference database. The results indicate a promising use for a SigClust-based pipeline in wound analysis and prediction, and a possible novel method for wound management and treatment.

Highlights

  • Sequencing highly-variable 16S regions is a common and often effective approach to the study of microbial communities, and next-generation sequencing (NGS) technologies provide abundant quantities of data for analysis

  • We report for each method the clustering time in minutes and the number of clusters returned

  • In this paper we have introduced SigClust, a novel, highspeed clustering approach which allows the accurate analysis of read collections at scale, potentially supporting the timely processing of clinical wound samples as part of an integrated pipeline

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Summary

Introduction

Sequencing highly-variable 16S regions is a common and often effective approach to the study of microbial communities, and next-generation sequencing (NGS) technologies provide abundant quantities of data for analysis. The impact of chronic wounds is expected to increase markedly as the population ages and as the incidence of type II diabetes increases in line with increased incidence of obesity It is well-established that bacterial populations in the wound may heavily influence the Chappell et al BMC Bioinformatics 2018, 19(Suppl 20):0. It will not be possible to deliver on this promise without methods capable of handling these large-scale datasets and rapidly identifying markers of healing or stagnation. Such algorithms will be able to predict the progression of wound conditions over time

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