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The oligonucleotide frequency derived error gradient and its application to the binning of metagenome fragments.

ABSTRACT: BACKGROUND: The characterisation, or binning, of metagenome fragments is an important first step to further downstream analysis of microbial consortia. Here, we propose a one-dimensional signature, OFDEG, derived from the oligonucleotide frequency profile of a DNA sequence, and show that it is possible to obtain a meaningful phylogenetic signal for relatively short DNA sequences. The one-dimensional signal is essentially a compact representation of higher dimensional feature spaces of greater complexity and is intended to improve on the tetranucleotide frequency feature space preferred by current compositional binning methods. RESULTS: We compare the fidelity of OFDEG against tetranucleotide frequency in both an unsupervised and semi-supervised setting on simulated metagenome benchmark data. Four tests were conducted using assembler output of Arachne and phrap, and for each, performance was evaluated on contigs which are greater than or equal to 8 kbp in length and contigs which are composed of at least 10 reads. Using both G-C content in conjunction with OFDEG gave an average accuracy of 96.75% (semi-supervised) and 95.19% (unsupervised), versus 94.25% (semi-supervised) and 82.35% (unsupervised) for tetranucleotide frequency. CONCLUSION: We have presented an observation of an alternative characteristic of DNA sequences. The proposed feature representation has proven to be more beneficial than the existing tetranucleotide frequency space to the metagenome binning problem. We do note, however, that our observation of OFDEG deserves further anlaysis and investigation. Unsupervised clustering revealed OFDEG related features performed better than standard tetranucleotide frequency in representing a relevant organism specific signal. Further improvement in binning accuracy is given by semi-supervised classification using OFDEG. The emphasis on a feature-driven, bottom-up approach to the problem of binning reveals promising avenues for future development of techniques to characterise short environmental sequences without bias toward cultivable organisms.

SUBMITTER: Saeed I

PROVIDER: S-EPMC2788362 | biostudies-literature | 2009

REPOSITORIES: biostudies-literature

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The oligonucleotide frequency derived error gradient and its application to the binning of metagenome fragments.

Saeed Isaam I Halgamuge Saman K SK

BMC genomics 20091203

<h4>Background</h4>The characterisation, or binning, of metagenome fragments is an important first step to further downstream analysis of microbial consortia. Here, we propose a one-dimensional signature, OFDEG, derived from the oligonucleotide frequency profile of a DNA sequence, and show that it is possible to obtain a meaningful phylogenetic signal for relatively short DNA sequences. The one-dimensional signal is essentially a compact representation of higher dimensional feature spaces of gre ...[more]

PMID: 19958473

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Project description:Metagenomics sequencing provides deep insights into microbial communities. To investigate their taxonomic structure, binning assembled contigs into discrete clusters is critical. Many binning algorithms have been developed, but their performance is not always satisfactory, especially for complex microbial communities, calling for further development.According to previous studies, relative sequence compositions are similar across different regions of the same genome, but they differ between distinct genomes. Generally, current tools have used the normalized frequency of k-tuples directly, but this represents an absolute, not relative, sequence composition. Therefore, we attempted to model contigs using relative k-tuple composition, followed by measuring dissimilarity between contigs using [Formula: see text]. The [Formula: see text] was designed to measure the dissimilarity between two long sequences or Next-Generation Sequencing data with the Markov models of the background genomes. This method was effective in revealing group and gradient relationships between genomes, metagenomes and metatranscriptomes. With many binning tools available, we do not try to bin contigs from scratch. Instead, we developed [Formula: see text] to adjust contigs among bins based on the output of existing binning tools for a single metagenomic sample. The tool is taxonomy-free and depends only on k-tuples. To evaluate the performance of [Formula: see text], five widely used binning tools with different strategies of sequence composition or the hybrid of sequence composition and abundance were selected to bin six synthetic and real datasets, after which [Formula: see text] was applied to adjust the binning results. Our experiments showed that [Formula: see text] consistently achieves the best performance with tuple length k?=?6 under the independent identically distributed (i.i.d.) background model. Using the metrics of recall, precision and ARI (Adjusted Rand Index), [Formula: see text] improves the binning performance in 28 out of 30 testing experiments (6 datasets with 5 binning tools). The [Formula: see text] is available at https://github.com/kunWangkun/d2SBin .Experiments showed that [Formula: see text] accurately measures the dissimilarity between contigs of metagenomic reads and that relative sequence composition is more reasonable to bin the contigs. The [Formula: see text] can be applied to any existing contig-binning tools for single metagenomic samples to obtain better binning results.

Project description:BackgroundIn metagenomic studies, a process called binning is necessary to assign contigs that belong to multiple species to their respective phylogenetic groups. Most of the current methods of binning, such as BLAST, k-mer and PhyloPythia, involve assigning sequence fragments by comparing sequence similarity or sequence composition with already-sequenced genomes that are still far from comprehensive. We propose a semi-supervised seeding method for binning that does not depend on knowledge of completed genomes. Instead, it extracts the flanking sequences of highly conserved 16S rRNA from the metagenome and uses them as seeds (labels) to assign other reads based on their compositional similarity.ResultsThe proposed seeding method is implemented on an unsupervised Growing Self-Organising Map (GSOM), and called Seeded GSOM (S-GSOM). We compared it with four well-known semi-supervised learning methods in a preliminary test, separating random-length prokaryotic sequence fragments sampled from the NCBI genome database. We identified the flanking sequences of the highly conserved 16S rRNA as suitable seeds that could be used to group the sequence fragments according to their species. S-GSOM showed superior performance compared to the semi-supervised methods tested. Additionally, S-GSOM may also be used to visually identify some species that do not have seeds. The proposed method was then applied to simulated metagenomic datasets using two different confidence threshold settings and compared with PhyloPythia, k-mer and BLAST. At the reference taxonomic level Order, S-GSOM outperformed all k-mer and BLAST results and showed comparable results with PhyloPythia for each of the corresponding confidence settings, where S-GSOM performed better than PhyloPythia in the >/= 10 reads datasets and comparable in the > or = 8 kb benchmark tests.ConclusionIn the task of binning using semi-supervised learning methods, results indicate S-GSOM to be the best of the methods tested. Most importantly, the proposed method does not require knowledge from known genomes and uses only very few labels (one per species is sufficient in most cases), which are extracted from the metagenome itself. These advantages make it a very attractive binning method. S-GSOM outperformed the binning methods that depend on already-sequenced genomes, and compares well to the current most advanced binning method, PhyloPythia.

Dataset Information

The oligonucleotide frequency derived error gradient and its application to the binning of metagenome fragments.

Publications

The oligonucleotide frequency derived error gradient and its application to the binning of metagenome fragments.

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