Monday, May 6, 2013

Three Metagenomics Papers for You

A handful of good metagenomics papers have come out over the last few months. Below I've linked to and copied my evaluation of each of these articles from F1000.

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1. Willner, Dana, and Philip Hugenholtz. "From deep sequencing to viral tagging: Recent advances in viral metagenomics." BioEssays (2013). 

My evaluation: This review lays out some of the challenges and recent advances in viral metagenomic sequencing. There is a good discussion of library preparation and how that affects downstream sequencing. Alarmingly, they reference another paper that showed that different amplification methods resulted in detection of a completely different set of viruses (dsDNA viruses with LASL, ssDNA with MDA). The review also discusses many of the data management, analysis, and bioinformatics challenges associated with viral metagenomics.

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2. Loman, Nicholas J., et al. "A Culture-Independent Sequence-Based Metagenomics Approach to the Investigation of an Outbreak of Shiga-Toxigenic Escherichia coli O104: H4Outbreak of Shiga-toxigenic Escherichia coli." JAMA 309.14 (2013): 1502-1510.

My evaluation: This paper is a groundbreaking exploration of the use of metagenomics to investigate and determine the causal organism of an infectious disease outbreak. The authors retrospectively collected fecal samples from symptomatic patients from the 2011 Escherichia coli O104:H4 outbreak in Germany and performed high-throughput shotgun sequencing, followed by a sophisticated analysis to determine the outbreak's causal organism. The analysis included comparing genetic markers from many symptomatic patients' metagenomes with those of healthy controls, followed by de novo assembly of the outbreak strain from the shotgun metagenomic data. This illustrates both the power, but the real limitations, of using metagenomic approaches for clinical diagnostics. Also see David Relman's synopsis of the study in the same JAMA issue

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3. Shakya, Migun, et al. "Comparative metagenomic and rRNA microbial diversity characterization using archaeal and bacterial synthetic communities." Environmental microbiology (2013).

My evaluation: This study set out to compare shotgun metagenomic sequencing to 16S rRNA amplicon sequencing to determine the taxonomic and abundance profiles of mixed community metagenomic samples. Thus far, benchmarking metagenomic methodology has been difficult due to the lack of datasets where the underlying ground truth is known. In this study, the researchers constructed synthetic metagenomic communities consisting of 64 laboratory mixed genome DNAs of known sequence and polymerase chain reaction (PCR)-validated abundance. The researchers then compared metagenomic and 16S amplicon sequencing, using both 454 and Illumina technology, and found that metagenomic sequencing outperformed 16S sequencing in quantifying community composition. The synthetic metagenomes constructed here are publicly available (Gene Expression Omnibus [GEO] accession numbers are given in the manuscript), which represent a great asset to other researchers developing methods for amplicon-based or metagenomic approaches to sequence classification, diversity analysis, and abundance estimation.

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