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'Oumuamua (1I/2017 U1) is the first known object of interstellar origin to have entered the Solar System on an unbound and hyperbolic trajectory with respect to the Sun1. Various physical observations collected during its visit to the Solar System showed that it has an unusually elongated shape and a tumbling rotation state1-4 and that the physical properties of its surface resemble those of cometary nuclei5,6, even though it showed no evidence of cometary activity1,5,7. The motion of all celestial bodies is governed mostly by gravity, but the trajectories of comets can also be affected by non-gravitational forces due to cometary outgassing8. Because non-gravitational accelerations are at least three to four orders of magnitude weaker than gravitational acceleration, the detection of any deviation from a purely gravity-driven trajectory requires high-quality astrometry over a long arc. As a result, non-gravitational effects have been measured on only a limited subset of the small-body population9. Here we report the detection, at 30σ significance, of non-gravitational acceleration in the motion of 'Oumuamua. We analyse imaging data from extensive observations by ground-based and orbiting facilities. This analysis rules out systematic biases and shows that all astrometric data can be described once a non-gravitational component representing a heliocentric radial acceleration proportional to r-2 or r-1 (where r is the heliocentric distance) is included in the model. After ruling out solar-radiation pressure, drag- and friction-like forces, interaction with solar wind for a highly magnetized object, and geometric effects originating from 'Oumuamua potentially being composed of several spatially separated bodies or having a pronounced offset between its photocentre and centre of mass, we find comet-like outgassing to be a physically viable explanation, provided that 'Oumuamua has thermal properties similar to comets.
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None of the approximately 750,000 known asteroids and comets in the Solar System is thought to have originated outside it, despite models of the formation of planetary systems suggesting that orbital migration of giant planets ejects a large fraction of the original planetesimals into interstellar space. The high predicted number density of icy interstellar objects (2.4 × 10-4 per cubic astronomical unit) suggests that some should have been detected, yet hitherto none has been seen. Many decades of asteroid and comet characterization have yielded formation models that explain the mass distribution, chemical abundances and planetary configuration of the Solar System today, but there has been no way of telling whether the Solar System is typical of planetary systems. Here we report observations and analysis of the object 1I/2017 U1 ('Oumuamua) that demonstrate its extrasolar trajectory, and that thus enable comparisons to be made between material from another planetary system and from our own. Our observations during the brief visit by the object to the inner Solar System reveal it to be asteroidal, with no hint of cometary activity despite an approach within 0.25 astronomical units of the Sun. Spectroscopic measurements show that the surface of the object is spectrally red, consistent with comets or organic-rich asteroids that reside within the Solar System. Light-curve observations indicate that the object has an extremely oblong shape, with a length about ten times its width, and a mean radius of about 102 metres assuming an albedo of 0.04. No known objects in the Solar System have such extreme dimensions. The presence of 'Oumuamua in the Solar System suggests that previous estimates of the number density of interstellar objects, based on the assumption that all such objects were cometary, were pessimistically low. Planned upgrades to contemporary asteroid survey instruments and improved data processing techniques are likely to result in the detection of more interstellar objects in the coming years.
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Tapeworms (Cestoda) cause neglected diseases that can be fatal and are difficult to treat, owing to inefficient drugs. Here we present an analysis of tapeworm genome sequences using the human-infective species Echinococcus multilocularis, E. granulosus, Taenia solium and the laboratory model Hymenolepis microstoma as examples. The 115- to 141-megabase genomes offer insights into the evolution of parasitism. Synteny is maintained with distantly related blood flukes but we find extreme losses of genes and pathways that are ubiquitous in other animals, including 34 homeobox families and several determinants of stem cell fate. Tapeworms have specialized detoxification pathways, metabolism that is finely tuned to rely on nutrients scavenged from their hosts, and species-specific expansions of non-canonical heat shock proteins and families of known antigens. We identify new potential drug targets, including some on which existing pharmaceuticals may act. The genomes provide a rich resource to underpin the development of urgently needed treatments and control.
Assuntos
Adaptação Fisiológica/genética , Cestoides/genética , Genoma Helmíntico/genética , Parasitos/genética , Animais , Evolução Biológica , Cestoides/efeitos dos fármacos , Cestoides/fisiologia , Infecções por Cestoides/tratamento farmacológico , Infecções por Cestoides/metabolismo , Sequência Conservada/genética , Echinococcus granulosus/genética , Echinococcus multilocularis/efeitos dos fármacos , Echinococcus multilocularis/genética , Echinococcus multilocularis/metabolismo , Genes de Helmintos/genética , Genes Homeobox/genética , Proteínas de Choque Térmico HSP70/genética , Humanos , Hymenolepis/genética , Redes e Vias Metabólicas/genética , Terapia de Alvo Molecular , Parasitos/efeitos dos fármacos , Parasitos/fisiologia , Proteoma/genética , Células-Tronco/citologia , Células-Tronco/metabolismo , Taenia solium/genéticaRESUMO
Klebsiella pneumoniae shows increasing emergence of multidrug-resistant lineages, including strains resistant to all available antimicrobial drugs. We conducted whole-genome sequencing of 178 highly drug-resistant isolates from a tertiary hospital in Lahore, Pakistan. Phylogenetic analyses to place these isolates into global context demonstrate the expansion of multiple independent lineages, including K. quasipneumoniae.
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Farmacorresistência Bacteriana Múltipla/genética , Genoma Bacteriano/genética , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , Adolescente , Antibacterianos/farmacologia , Criança , Criança Hospitalizada , Pré-Escolar , Humanos , Lactente , Recém-Nascido , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/isolamento & purificação , Paquistão/epidemiologia , Filogenia , Análise de Sequência de DNARESUMO
UNLABELLED: Transposon insertion sequencing is a high-throughput technique for assaying large libraries of otherwise isogenic transposon mutants providing insight into gene essentiality, gene function and genetic interactions. We previously developed the Transposon Directed Insertion Sequencing (TraDIS) protocol for this purpose, which utilizes shearing of genomic DNA followed by specific PCR amplification of transposon-containing fragments and Illumina sequencing. Here we describe an optimized high-yield library preparation and sequencing protocol for TraDIS experiments and a novel software pipeline for analysis of the resulting data. The Bio-Tradis analysis pipeline is implemented as an extensible Perl library which can either be used as is, or as a basis for the development of more advanced analysis tools. This article can serve as a general reference for the application of the TraDIS methodology. AVAILABILITY AND IMPLEMENTATION: The optimized sequencing protocol is included as supplementary information. The Bio-Tradis analysis pipeline is available under a GPL license at https://github.com/sanger-pathogens/Bio-Tradis CONTACT: parkhill@sanger.ac.uk SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
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Elementos de DNA Transponíveis , Biblioteca Gênica , Software , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
The emergence of new sequencing technologies has facilitated the use of bacterial whole genome alignments for evolutionary studies and outbreak analyses. These datasets, of increasing size, often include examples of multiple different mechanisms of horizontal sequence transfer resulting in substantial alterations to prokaryotic chromosomes. The impact of these processes demands rapid and flexible approaches able to account for recombination when reconstructing isolates' recent diversification. Gubbins is an iterative algorithm that uses spatial scanning statistics to identify loci containing elevated densities of base substitutions suggestive of horizontal sequence transfer while concurrently constructing a maximum likelihood phylogeny based on the putative point mutations outside these regions of high sequence diversity. Simulations demonstrate the algorithm generates highly accurate reconstructions under realistically parameterized models of bacterial evolution, and achieves convergence in only a few hours on alignments of hundreds of bacterial genome sequences. Gubbins is appropriate for reconstructing the recent evolutionary history of a variety of haploid genotype alignments, as it makes no assumptions about the underlying mechanism of recombination. The software is freely available for download at github.com/sanger-pathogens/Gubbins, implemented in Python and C and supported on Linux and Mac OS X.
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Algoritmos , Bactérias/classificação , Genoma Bacteriano , Filogenia , Recombinação Genética , Bactérias/genética , Análise de SequênciaRESUMO
MOTIVATION: An accurate genome assembly from short read sequencing data is critical for downstream analysis, for example allowing investigation of variants within a sequenced population. However, assembling sequencing data from virus samples, especially RNA viruses, into a genome sequence is challenging due to the combination of viral population diversity and extremely uneven read depth caused by amplification bias in the inevitable reverse transcription and polymerase chain reaction amplification process of current methods. RESULTS: We developed a new de novo assembler called IVA (Iterative Virus Assembler) designed specifically for read pairs sequenced at highly variable depth from RNA virus samples. We tested IVA on datasets from 140 sequenced samples from human immunodeficiency virus-1 or influenza-virus-infected people and demonstrated that IVA outperforms all other virus de novo assemblers. AVAILABILITY AND IMPLEMENTATION: The software runs under Linux, has the GPLv3 licence and is freely available from http://sanger-pathogens.github.io/iva
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Genoma Viral , HIV-1/genética , Vírus da Influenza A/genética , Vírus da Influenza B/genética , Vírus de RNA/genética , Análise de Sequência de DNA/métodos , Software , Infecções por HIV/genética , Infecções por HIV/virologia , HIV-1/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Vírus da Influenza A/isolamento & purificação , Vírus da Influenza B/isolamento & purificação , Influenza Humana/genética , Influenza Humana/virologia , Reação em Cadeia da Polimerase/métodosRESUMO
UNLABELLED: A typical prokaryote population sequencing study can now consist of hundreds or thousands of isolates. Interrogating these datasets can provide detailed insights into the genetic structure of prokaryotic genomes. We introduce Roary, a tool that rapidly builds large-scale pan genomes, identifying the core and accessory genes. Roary makes construction of the pan genome of thousands of prokaryote samples possible on a standard desktop without compromising on the accuracy of results. Using a single CPU Roary can produce a pan genome consisting of 1000 isolates in 4.5 hours using 13 GB of RAM, with further speedups possible using multiple processors. AVAILABILITY AND IMPLEMENTATION: Roary is implemented in Perl and is freely available under an open source GPLv3 license from http://sanger-pathogens.github.io/Roary CONTACT: roary@sanger.ac.uk SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
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Genoma Bacteriano , Células Procarióticas/metabolismo , Software , Simulação por Computador , Bases de Dados Genéticas , Salmonella typhi/genéticaRESUMO
Background. PCR amplification is a necessary step in many next-generation sequencing (NGS) library preparation methods [1, 2]. Whilst many PCR enzymes are developed to amplify single targets efficiently, accurately and with specificity, few are developed to meet the challenges imposed by NGS PCR, namely unbiased amplification of a wide range of different sizes and GC content. As a result PCR amplification during NGS library prep often results in bias toward GC neutral and smaller fragments. As NGS has matured, optimized NGS library prep kits and polymerase formulations have emerged and in this study we have tested a wide selection of available enzymes for both short-read Illumina library preparation and long fragment amplification ahead of long-read sequencing.We tested over 20 different hi-fidelity PCR enzymes/NGS amplification mixes on a range of Illumina library templates of varying GC content and composition, and find that both yield and genome coverage uniformity characteristics of the commercially available enzymes varied dramatically. Three enzymes Quantabio RepliQa Hifi Toughmix, Watchmaker Library Amplification Hot Start Master Mix (2X) 'Equinox' and Takara Ex Premier were found to give a consistent performance, over all genomes, that mirrored closely that observed for PCR-free datasets. We also test a range of enzymes for long-read sequencing by amplifying size fractionated S. cerevisiae DNA of average size 21.6 and 13.4 kb, respectively.The enzymes of choice for short-read (Illumina) library fragment amplification are Quantabio RepliQa Hifi Toughmix, Watchmaker Library Amplification Hot Start Master Mix (2X) 'Equinox' and Takara Ex Premier, with RepliQa also being the best performing enzyme from the enzymes tested for long fragment amplification prior to long-read sequencing.
Assuntos
DNA , Saccharomyces cerevisiae , Reação em Cadeia da Polimerase/métodos , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala/métodosRESUMO
Antimicrobial resistance (AMR) poses a serious threat to the clinical management of typhoid fever. AMR in Salmonella Typhi (S. Typhi) is commonly associated with the H58 lineage, a lineage that arose comparatively recently before becoming globally disseminated. To better understand when and how H58 emerged and became dominant, we performed detailed phylogenetic analyses on contemporary genome sequences from S. Typhi isolated in the period spanning the emergence. Our dataset, which contains the earliest described H58 S. Typhi organism, indicates that ancestral H58 organisms were already multi-drug resistant (MDR). These organisms emerged spontaneously in India in 1987 and became radially distributed throughout South Asia and then globally in the ensuing years. These early organisms were associated with a single long branch, possessing mutations associated with increased bile tolerance, suggesting that the first H58 organism was generated during chronic carriage. The subsequent use of fluoroquinolones led to several independent mutations in gyrA. The ability of H58 to acquire and maintain AMR genes continues to pose a threat, as extensively drug-resistant (XDR; MDR plus resistance to ciprofloxacin and third generation cephalosporins) variants, have emerged recently in this lineage. Understanding where and how H58 S. Typhi originated and became successful is key to understand how AMR drives successful lineages of bacterial pathogens. Additionally, these data can inform optimal targeting of typhoid conjugate vaccines (TCVs) for reducing the potential for emergence and the impact of new drug-resistant variants. Emphasis should also be placed upon the prospective identification and treatment of chronic carriers to prevent the emergence of new drug resistant variants with the ability to spread efficiently.
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Antibacterianos , Filogenia , Salmonella typhi , Febre Tifoide , Salmonella typhi/genética , Salmonella typhi/efeitos dos fármacos , Febre Tifoide/microbiologia , Febre Tifoide/tratamento farmacológico , Febre Tifoide/epidemiologia , Humanos , Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Haplótipos , Mutação , Genoma BacterianoRESUMO
Salmonella Paratyphi A, one of the major etiologic agents of enteric fever, has increased in prevalence in recent decades in certain endemic regions in comparison to S. Typhi, the most prevalent cause of enteric fever. Despite this increase, data on the prevalence and molecular epidemiology of S. Paratyphi A remain generally scarce. Here, we analysed the whole genome sequences of 216 S. Paratyphi A isolates originating from Kathmandu, Nepal between 2005 and 2014, of which 200 were from patients with acute enteric fever and 16 from the gallbladder of people with suspected chronic carriage. By exploiting the recently developed genotyping framework for S. Paratyphi A (Paratype), we identified several genotypes circulating in Kathmandu. Notably, we observed an unusual clonal expansion of genotype 2.4.3 over a four-year period that spread geographically and systematically replaced other genotypes. This rapid genotype replacement is hypothesised to have been driven by both reduced susceptibility to fluoroquinolones and genetic changes to virulence factors, such as functional and structural genes encoding the type 3 secretion systems. Finally, we show that person-to-person is likely the most common mode of transmission and chronic carriers seem to play a limited role in maintaining disease circulation.
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Genótipo , Febre Paratifoide , Salmonella paratyphi A , Nepal/epidemiologia , Humanos , Salmonella paratyphi A/genética , Salmonella paratyphi A/isolamento & purificação , Salmonella paratyphi A/classificação , Estudos Retrospectivos , Febre Paratifoide/epidemiologia , Febre Paratifoide/microbiologia , Masculino , Adulto , Feminino , Adulto Jovem , Adolescente , Criança , Prevalência , Pessoa de Meia-Idade , Epidemiologia Molecular , Pré-Escolar , Sequenciamento Completo do Genoma , Antibacterianos/farmacologia , FilogeniaRESUMO
Two-thirds of small-bowel transplantation (SBT) recipients develop bacteremia, with the majority of infections occurring within 3 months post-transplant. Sepsis-related mortality occurs in 31% of patients and is commonly caused by bacteria of gut origin, which are thought to translocate across the implanted organ. Serial post-transplant surveillance endoscopies provide an opportunity to study whether the composition of the ileal and colonic microbiota can predict the emergence as well as the pathogen of subsequent clinical infections in the SBT patient population. Five participants serially underwent aspiration of ileal and colonic bowel effluents at transplantation and during follow-up endoscopy either until death or for up to 3 months post-SBT. We performed whole-metagenome sequencing (WMS) of 40 bowel effluent samples and compared the results with clinical infection episodes. Microbiome composition was concordant between participants and timepoint-matched ileal and colonic samples. Four out of five (4/5) participants had clinically significant infections thought to be of gut origin. Bacterial translocation from the gut was observed in 3/5 patients with bacterial infectious etiologies. In all three cases, the pathogens had demonstrably colonized the gut between 1-10 days prior to invasive clinical infection. Recipients with better outcomes received donor grafts with higher alpha diversity. There was an increase in the number of antimicrobial resistance genes associated with longer hospital stay for all participants. This metagenomic study provides preliminary evidence to support the pathogen translocation hypothesis of gut-origin sepsis in the SBT cohort. Ileal and colonic microbiome compositions were concordant; therefore, fecal metagenomic analysis could be a useful surveillance tool for impeding infection with specific gut-residing pathogens.
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Microbioma Gastrointestinal , Microbiota , Sepse , Humanos , Microbioma Gastrointestinal/genética , Metagenoma , Estudos ProspectivosRESUMO
The National Collection of Type Cultures (NCTC) was founded on 1 January 1920 in order to fulfil a recognized need for a centralized repository for bacterial and fungal strains within the UK. It is among the longest-established collections of its kind anywhere in the world and today holds approximately 6000 type and reference bacterial strains - many of medical, scientific and veterinary importance - available to academic, health, food and veterinary institutions worldwide. Recently, a collaboration between NCTC, Pacific Biosciences and the Wellcome Sanger Institute established the NCTC3000 project to long-read sequence and assemble the genomes of up to 3000 NCTC strains. Here, at the beginning of the collection's second century, we introduce the resulting NCTC3000 sequence read datasets, genome assemblies and annotations as a unique, historically and scientifically relevant resource for the benefit of the international bacterial research community.
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Genoma Bacteriano , Genômica , Análise de Sequência de DNA/métodos , Genoma Bacteriano/genética , Bactérias/genéticaRESUMO
As whole-genome sequencing capacity becomes increasingly decentralized, there is a growing opportunity for collaboration and the sharing of surveillance data within and between countries to inform typhoid control policies. This vision requires free, community-driven tools that facilitate access to genomic data for public health on a global scale. Here we present the Pathogenwatch scheme for Salmonella enterica serovar Typhi (S. Typhi), a web application enabling the rapid identification of genomic markers of antimicrobial resistance (AMR) and contextualization with public genomic data. We show that the clustering of S. Typhi genomes in Pathogenwatch is comparable to established bioinformatics methods, and that genomic predictions of AMR are highly concordant with phenotypic susceptibility data. We demonstrate the public health utility of Pathogenwatch with examples selected from >4,300 public genomes available in the application. Pathogenwatch provides an intuitive entry point to monitor of the emergence and spread of S. Typhi high risk clones.
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Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Salmonella typhi/efeitos dos fármacos , Febre Tifoide/prevenção & controle , Proteínas de Bactérias/genética , Genoma Bacteriano/genética , Genômica/métodos , Genótipo , Geografia , Humanos , Malaui , Proteínas de Membrana Transportadoras/genética , Testes de Sensibilidade Microbiana/métodos , Mutação , Salmonella typhi/genética , Salmonella typhi/fisiologia , Tanzânia , Febre Tifoide/microbiologiaRESUMO
Schistosomes are parasitic blood flukes that survive for many years within the mammalian host vasculature. How the parasites establish a chronic infection in the hostile bloodstream environment, whilst evading the host immune response is poorly understood. The parasite develops morphologically and grows as it migrates to its preferred vascular niche, avoiding or repairing damage from the host immune system. In this study, we investigated temporal changes in gene expression during the intra-mammalian development of Schistosoma mansoni. RNA-seq data were analysed from parasites developing in the lung through to egg-laying mature adult worms, providing a comprehensive picture of in vivo intra-mammalian development. Remarkably, genes involved in signalling pathways, developmental control, and adaptation to oxidative stress were up-regulated in the lung stage. The data also suggested a potential role in immune evasion for a previously uncharacterised gene. This study not only provides a large and comprehensive data resource for the research community, but also reveals new directions for further characterising host-parasite interactions that could ultimately lead to new control strategies for this neglected tropical disease pathogen.
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Proteínas de Helminto/genética , Schistosoma mansoni/crescimento & desenvolvimento , Schistosoma mansoni/genética , Esquistossomose mansoni/parasitologia , Animais , Feminino , Proteínas de Helminto/metabolismo , Humanos , Masculino , Camundongos , Schistosoma mansoni/metabolismo , Esquistossomose mansoni/patologia , TranscriptomaRESUMO
Genome sequencing is rapidly being adopted in reference labs and hospitals for bacterial outbreak investigation and diagnostics where time is critical. Seven gene multi-locus sequence typing is a standard tool for broadly classifying samples into sequence types (STs), allowing, in many cases, to rule a sample out of an outbreak, or allowing for general characteristics about a bacterial strain to be inferred. Long-read sequencing technologies, such as from Oxford Nanopore, can produce read data within minutes of an experiment starting, unlike short-read sequencing technologies which require many hours/days. However, the error rates of raw uncorrected long read data are very high. We present Krocus which can predict a ST directly from uncorrected long reads, and which was designed to consume read data as it is produced, providing results in minutes. It is the only tool which can do this from uncorrected long reads. We tested Krocus on over 700 isolates sequenced using long-read sequencing technologies from Pacific Biosciences and Oxford Nanopore. It provides STs for isolates on average within 90 s, with a sensitivity of 94% and specificity of 97% on real sample data, directly from uncorrected raw sequence reads. The software is written in Python and is available under the open source license GNU GPL version 3.
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Streptococcus pneumoniae is responsible for 240â000-460â000 deaths in children under 5 years of age each year. Accurate identification of pneumococcal serotypes is important for tracking the distribution and evolution of serotypes following the introduction of effective vaccines. Recent efforts have been made to infer serotypes directly from genomic data but current software approaches are limited and do not scale well. Here, we introduce a novel method, SeroBA, which uses a k-mer approach. We compare SeroBA against real and simulated data and present results on the concordance and computational performance against a validation dataset, the robustness and scalability when analysing a large dataset, and the impact of varying the depth of coverage on sequence-based serotyping. SeroBA can predict serotypes, by identifying the cps locus, directly from raw whole genome sequencing read data with 98â% concordance using a k-mer-based method, can process 10â000 samples in just over 1 day using a standard server and can call serotypes at a coverage as low as 15-21×. SeroBA is implemented in Python3 and is freely available under an open source GPLv3 licence from: https://github.com/sanger-pathogens/seroba.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Infecções Pneumocócicas/microbiologia , Sorotipagem/métodos , Software , Streptococcus mitis/genética , Streptococcus pneumoniae/classificação , Sequenciamento Completo do Genoma , Alelos , Pré-Escolar , Bases de Dados Genéticas , Genes Bacterianos , Humanos , Polimorfismo de Nucleotídeo Único , Sensibilidade e Especificidade , Sorogrupo , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/isolamento & purificaçãoRESUMO
Increasingly rich metadata are now being linked to samples that have been whole-genome sequenced. However, much of this information is ignored. This is because linking this metadata to genes, or regions of the genome, usually relies on knowing the gene sequence(s) responsible for the particular trait being measured and looking for its presence or absence in that genome. Examples of this would be the spread of antimicrobial resistance genes carried on mobile genetic elements (MGEs). However, although it is possible to routinely identify the resistance gene, identifying the unknown MGE upon which it is carried can be much more difficult if the starting point is short-read whole-genome sequence data. The reason for this is that MGEs are often full of repeats and so assemble poorly, leading to fragmented consensus sequences. Since mobile DNA, which can carry many clinically and ecologically important genes, has a different evolutionary history from the host, its distribution across the host population will, by definition, be independent of the host phylogeny. It is possible to use this phenomenon in a genome-wide association study to identify both the genes associated with the specific trait and also the DNA linked to that gene, for example the flanking sequence of the plasmid vector on which it is encoded, which follows the same patterns of distribution as the marker gene/sequence itself. We present PlasmidTron, which utilizes the phenotypic data normally available in bacterial population studies, such as antibiograms, virulence factors, or geographical information, to identify traits that are likely to be present on DNA that can randomly reassort across defined bacterial populations. It is also possible to use this methodology to associate unknown genes/sequences (e.g. plasmid backbones) with a specific molecular signature or marker (e.g. resistance gene presence or absence) using PlasmidTron. PlasmidTron uses a k-mer-based approach to identify reads associated with a phylogenetically unlinked phenotype. These reads are then assembled de novo to produce contigs in a fast and scalable-to-large manner. PlasmidTron is written in Python 3 and is available under the open source licence GNU GPL3 from https://github.com/sanger-pathogens/plasmidtron.
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Estudos de Associação Genética , Variações do Número de Cópias de DNA , Genoma Bacteriano , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/isolamento & purificação , Testes de Sensibilidade Microbiana , Fenótipo , Filogenia , Plasmídeos/genética , Plasmídeos/isolamento & purificação , Salmonella enterica/genética , Salmonella enterica/isolamento & purificação , Análise de Sequência de DNARESUMO
Multi-locus sequence typing (MLST) is a widely used method for categorizing bacteria. Increasingly, MLST is being performed using next-generation sequencing (NGS) data by reference laboratories and for clinical diagnostics. Many software applications have been developed to calculate sequence types from NGS data; however, there has been no comprehensive review to date on these methods. We have compared eight of these applications against real and simulated data, and present results on: (1) the accuracy of each method against traditional typing methods, (2) the performance on real outbreak datasets, (3) the impact of contamination and varying depth of coverage, and (4) the computational resource requirements.
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Bactérias/genética , Técnicas de Tipagem Bacteriana/métodos , Tipagem de Sequências Multilocus/métodos , Bases de Dados Factuais , Genoma Bacteriano , SoftwareRESUMO
Antimicrobial resistance (AMR) is one of the major threats to human and animal health worldwide, yet few high-throughput tools exist to analyse and predict the resistance of a bacterial isolate from sequencing data. Here we present a new tool, ARIBA, that identifies AMR-associated genes and single nucleotide polymorphisms directly from short reads, and generates detailed and customizable output. The accuracy and advantages of ARIBA over other tools are demonstrated on three datasets from Gram-positive and Gram-negative bacteria, with ARIBA outperforming existing methods.