Nanopore metagenomic sequencing of full length human metapneumovirus (HMPV) within a unique sub-lineage

Mapping Intimacies ◽

10.1101/496687 ◽

2018 ◽

Cited By ~ 2

Author(s):

Yifei Xu ◽

Kuiama Lewandowski ◽

Sheila Lumley ◽

Nicholas D Sanderson ◽

Alison Vaughan ◽

...

Keyword(s):

Phylogenetic Analyses ◽

Human Metapneumovirus ◽

Full Length ◽

Clinical Settings ◽

Metagenomic Sequencing ◽

Sequencing Data ◽

Complete Sequences ◽

Genomic Characterization ◽

Tract Disease ◽

The Uk

Human metapneumovirus (HMPV) has been recognized as an important pathogen which can cause a spectrum of respiratory tract disease. Here, we report Nanopore metagenomic sequencing of the first full length HMPV genome directly from a throat swab from a UK patient with complex lung disease and immunocompromise. We found a predominance (26.4%) of HMPV reads in the metagenomic sequencing data and consequently assembled the full genome at a high depth of coverage (mean 4,786). Through phylogenetic analyses, we identified this HMPV strain to originate from a unique genetic group in A2b, showing the presence of this group in the UK. Our study demonstrated the effectiveness of Nanopore metagenomic sequencing for diagnosing infectious diseases and recovering complete sequences for genomic characterization, highlighting the applicability of Nanopore sequencing in clinical settings.

Characterizing and Evaluating the Zoonotic Potential of Novel Viruses Discovered in Vampire Bats

Viruses ◽

10.3390/v13020252 ◽

2021 ◽

Vol 13 (2) ◽

pp. 252

Author(s):

Laura M. Bergner ◽

Nardus Mollentze ◽

Richard J. Orton ◽

Carlos Tello ◽

Alice Broos ◽

...

Keyword(s):

Machine Learning ◽

Phylogenetic Analyses ◽

Human Infection ◽

Machine Learning Algorithms ◽

Zoonotic Potential ◽

Metagenomic Sequencing ◽

Learning Models ◽

Sequencing Data ◽

Vampire Bats ◽

Machine Learning Models

The contemporary surge in metagenomic sequencing has transformed knowledge of viral diversity in wildlife. However, evaluating which newly discovered viruses pose sufficient risk of infecting humans to merit detailed laboratory characterization and surveillance remains largely speculative. Machine learning algorithms have been developed to address this imbalance by ranking the relative likelihood of human infection based on viral genome sequences, but are not yet routinely applied to viruses at the time of their discovery. Here, we characterized viral genomes detected through metagenomic sequencing of feces and saliva from common vampire bats (Desmodus rotundus) and used these data as a case study in evaluating zoonotic potential using molecular sequencing data. Of 58 detected viral families, including 17 which infect mammals, the only known zoonosis detected was rabies virus; however, additional genomes were detected from the families Hepeviridae, Coronaviridae, Reoviridae, Astroviridae and Picornaviridae, all of which contain human-infecting species. In phylogenetic analyses, novel vampire bat viruses most frequently grouped with other bat viruses that are not currently known to infect humans. In agreement, machine learning models built from only phylogenetic information ranked all novel viruses similarly, yielding little insight into zoonotic potential. In contrast, genome composition-based machine learning models estimated different levels of zoonotic potential, even for closely related viruses, categorizing one out of four detected hepeviruses and two out of three picornaviruses as having high priority for further research. We highlight the value of evaluating zoonotic potential beyond ad hoc consideration of phylogeny and provide surveillance recommendations for novel viruses in a wildlife host which has frequent contact with humans and domestic animals.

Detection of viral pathogens with multiplex Nanopore MinION sequencing: be careful with cross-talk

10.1101/308262 ◽

2018 ◽

Author(s):

Yifei Xu ◽

Kuiama Lewandowski ◽

Sheila Lumley ◽

Steven Pullan ◽

Richard Vipond ◽

...

Keyword(s):

Influenza A ◽

Point Of Care ◽

Clinical Settings ◽

Metagenomic Sequencing ◽

Sequencing Data ◽

Viral Pathogens ◽

Oxford Nanopore ◽

Downstream Analysis ◽

Multiplex Sequencing ◽

Single Flow

AbstractMetagenomic sequencing with the Oxford Nanopore MinION sequencer offers potential for point-of-care testing of infectious diseases in clinical settings. To improve cost-effectiveness, multiplexing of several, barcoded samples upon a single flow cell will be required during sequencing. We generated a unique sequencing dataset to assess the extent and source of cross barcode contamination caused by multiplex MinION sequencing. Sequencing libraries for three different viruses, including influenza A, dengue and chikungunya, were prepared separately and sequenced on individual flow cells. We also pooled the respective libraries and performed multiplex sequencing. We identified 0.056% of total reads in the multiplex sequencing data that were assigned to incorrect barcodes. Chimeric reads were the predominant source of this error. Our findings highlight the need for careful filtering of multiplex sequencing data before downstream analysis, and the trade-off between sensitivity and specificity that applies to the barcode demultiplexing methods.

Isolation and full genomic characterization of Batai virus from mosquitoes, Italy 2009

Journal of General Virology ◽

10.1099/vir.0.051359-0 ◽

2013 ◽

Vol 94 (6) ◽

pp. 1242-1248 ◽

Cited By ~ 18

Author(s):

Eili Huhtamo ◽

Amy J. Lambert ◽

Stefano Costantino ◽

Luca Servino ◽

Letizia Krizmancic ◽

...

Keyword(s):

Geographical Distribution ◽

Phylogenetic Analyses ◽

Full Length ◽

Limited Data ◽

Rt Pcr ◽

Anopheles Maculipennis ◽

Genomic Characterization ◽

Batai Virus

In 2009, 2589 mosquitoes were collected in northwest Italy and screened for orthobunyavirus RNA by RT-PCR. One pool of Anopheles maculipennis complex mosquitoes was found to be positive and a virus was isolated from that pool. The isolate was identified as Batai virus (BATV) by sequencing. Previously, BATV was detected in Italy, but limited data and no prior isolates existed. Full-length sequences of the S, M and L segments were determined for the newly isolated Italian strain. For comparison, partial sequences were also determined for the BATV strain Calovo (former Czechoslovakia, 1960). Phylogenetic analyses revealed clustering of the newly derived Italian BATV along with a recent isolate from Germany and the historic strain Calovo. To the best of our knowledge, this represents the first isolation of BATV from Italy, which confirms a broader geographical distribution of BATV in Europe than was previously verified by isolation.

Full-length genome sequence of segmented RNA virus from ticks was obtained using small RNA sequencing data

10.1101/2020.03.24.004796 ◽

2020 ◽

Author(s):

Xiaofeng Xu ◽

Jinlong Bei ◽

Yibo Xuan ◽

Jianyuan Chen ◽

Defu Chen ◽

...

Keyword(s):

Rna Sequencing ◽

Genome Sequence ◽

Small Rna ◽

Rna Virus ◽

Phylogenetic Analyses ◽

Full Length ◽

Small Rna Sequencing ◽

Sequencing Data ◽

Link Type ◽

Full Length Genome

AbstractIn 2014, A novel tick-borne virus of the genus Flavivirus was first reported from the Mogiana region in Brazil. This virus was named the Mogiana tick virus (MGTV). Later, MGTV was also named as Jingmen tick virus (JMTV), Kindia tick virus (KDTV), Guangxi tick virus (GXTV) etc. In the present study, we used small RNA sequencing (sRNA-seq) to detect viruses in ticks and detected MGTV in Amblyomma testudinarium ticks, which had been captured in Yunnan province of China in the year of 2016. The full-length genome sequence of a new MGTV strain Yunnan2016 (GenBank: MT080097, MT080098, MT080099 and MT080100) was obtained and recommended to be included into the NCBI RefSeq database for the future studies of MGTV. Our phylogenetic analyses showed that viruses named MGTV, JMTV, KDTV and GXTV are monophyletic: the MGTV group (lineage) of viruses. We show, for the first time, that 5′ and 3′ sRNAs can be used to obtain full-length sequences of the 5’ and 3’ ends of, but not limited to genome sequences of RNA viruses. And we proved the feasibility of using the sRNA-seq based method for the detection of viruses in a sample containing miniscule RNA.

Use of SNP chips to detect rare pathogenic variants: retrospective, population based diagnostic evaluation

BMJ ◽

10.1136/bmj.n214 ◽

2021 ◽

pp. n214

Author(s):

Weedon MN ◽

Jackson L ◽

Harrison JW ◽

Ruth KS ◽

Tyrrell J ◽

...

Keyword(s):

Positive Predictive Value ◽

Predictive Value ◽

Population Based ◽

Genome Project ◽

Personal Genome ◽

Uk Biobank ◽

Sequencing Data ◽

Snp Chip ◽

Pathogenic Variants ◽

The Uk

Abstract Objective To determine whether the sensitivity and specificity of SNP chips are adequate for detecting rare pathogenic variants in a clinically unselected population. Design Retrospective, population based diagnostic evaluation. Participants 49 908 people recruited to the UK Biobank with SNP chip and next generation sequencing data, and an additional 21 people who purchased consumer genetic tests and shared their data online via the Personal Genome Project. Main outcome measures Genotyping (that is, identification of the correct DNA base at a specific genomic location) using SNP chips versus sequencing, with results split by frequency of that genotype in the population. Rare pathogenic variants in the BRCA1 and BRCA2 genes were selected as an exemplar for detailed analysis of clinically actionable variants in the UK Biobank, and BRCA related cancers (breast, ovarian, prostate, and pancreatic) were assessed in participants through use of cancer registry data. Results Overall, genotyping using SNP chips performed well compared with sequencing; sensitivity, specificity, positive predictive value, and negative predictive value were all above 99% for 108 574 common variants directly genotyped on the SNP chips and sequenced in the UK Biobank. However, the likelihood of a true positive result decreased dramatically with decreasing variant frequency; for variants that are very rare in the population, with a frequency below 0.001% in UK Biobank, the positive predictive value was very low and only 16% of 4757 heterozygous genotypes from the SNP chips were confirmed with sequencing data. Results were similar for SNP chip data from the Personal Genome Project, and 20/21 individuals analysed had at least one false positive rare pathogenic variant that had been incorrectly genotyped. For pathogenic variants in the BRCA1 and BRCA2 genes, which are individually very rare, the overall performance metrics for the SNP chips versus sequencing in the UK Biobank were: sensitivity 34.6%, specificity 98.3%, positive predictive value 4.2%, and negative predictive value 99.9%. Rates of BRCA related cancers in UK Biobank participants with a positive SNP chip result were similar to those for age matched controls (odds ratio 1.31, 95% confidence interval 0.99 to 1.71) because the vast majority of variants were false positives, whereas sequence positive participants had a significantly increased risk (odds ratio 4.05, 2.72 to 6.03). Conclusions SNP chips are extremely unreliable for genotyping very rare pathogenic variants and should not be used to guide health decisions without validation.

A Delphi Study Investigating Clinicians’ Views on Access to, Delivery of, and Adaptations of MBCT in the UK Clinical Settings

Mindfulness ◽

10.1007/s12671-021-01706-5 ◽

2021 ◽

Author(s):

Kate Williams ◽

Samantha Hartley ◽

Peter Taylor

Keyword(s):

Delphi Study ◽

Good Practice ◽

Evidence Base ◽

Clinical Settings ◽

Training Support ◽

Decision Making Processes ◽

Complex Decision Making ◽

Complex Decision ◽

The Uk ◽

Clinical Populations

Abstract Objectives Mindfulness-based cognitive therapy (MBCT) is a well-evidenced relapse-prevention intervention for depression with a growing evidence-base for use in other clinical populations. The UK initiatives have outlined plans for increasing access to MBCT in clinical settings, although evidence suggests that access remains limited. Given the increased popularity and access to MBCT, there may be deviations from the evidence-base and potential risks of harm. We aimed to understand what clinicians believe should be best clinical practice regarding access to, delivery of, and adaptations to MBCT. Methods We employed a two-stage Delphi methodology. First, to develop statements around best practices, we consulted five mindfulness-based experts and reviewed the literature. Second, a total of 59 statements were taken forward into three survey rating rounds. Results Twenty-nine clinicians completed round one, with 25 subsequently completing both rounds two and three. Forty-four statements reached consensus; 15 statements did not. Clinicians agreed with statements regarding sufficient preparation for accessing MBCT, adherence to the evidence-base and good practice guidelines, consideration of risks, sufficient access to training, support, and resources within services, and carefully considered adaptations. The consensus was not reached on statements which reflected a lack of evidence-base for specific clinical populations or the complex decision-making processes involved in delivering and making adaptations to MBCT. Conclusions Our findings highlight the delicate balance of maintaining a client-centred and transparent approach whilst adhering to the evidence-base in clinical decisions around access to, delivery of, and adaptations in MBCT and have important wide-reaching implications.

Establishment and lineage dynamics of the SARS-CoV-2 epidemic in the UK

Science ◽

10.1126/science.abf2946 ◽

2021 ◽

pp. eabf2946

Author(s):

Louis du Plessis ◽

John T. McCrone ◽

Alexander E. Zarebski ◽

Verity Hill ◽

Christopher Ruis ◽

...

Keyword(s):

Large Scale ◽

Phylogenetic Analyses ◽

Transmission Dynamics ◽

Genetic Lineage ◽

Size Dependent ◽

Spatio Temporal ◽

The Uk ◽

Lineage Structure ◽

And Control ◽

Lineage Diversity

The UK’s COVID-19 epidemic during early 2020 was one of world’s largest and unusually well represented by virus genomic sampling. Here we reveal the fine-scale genetic lineage structure of this epidemic through analysis of 50,887 SARS-CoV-2 genomes, including 26,181 from the UK sampled throughout the country’s first wave of infection. Using large-scale phylogenetic analyses, combined with epidemiological and travel data, we quantify the size, spatio-temporal origins and persistence of genetically-distinct UK transmission lineages. Rapid fluctuations in virus importation rates resulted in >1000 lineages; those introduced prior to national lockdown tended to be larger and more dispersed. Lineage importation and regional lineage diversity declined after lockdown, while lineage elimination was size-dependent. We discuss the implications of our genetic perspective on transmission dynamics for COVID-19 epidemiology and control.

METAMVGL: a multi-view graph-based metagenomic contig binning algorithm by integrating assembly and paired-end graphs

BMC Bioinformatics ◽

10.1186/s12859-021-04284-4 ◽

2021 ◽

Vol 22 (S10) ◽

Author(s):

Zhenmiao Zhang ◽

Lu Zhang

Keyword(s):

De Novo ◽

Label Propagation ◽

Next Generation Sequencing Data ◽

Metagenomic Sequencing ◽

Sequencing Data ◽

Fecal Samples ◽

Microbial Genomes ◽

Metagenome Assembly ◽

High Chance ◽

Mock Communities

Abstract Background Due to the complexity of microbial communities, de novo assembly on next generation sequencing data is commonly unable to produce complete microbial genomes. Metagenome assembly binning becomes an essential step that could group the fragmented contigs into clusters to represent microbial genomes based on contigs’ nucleotide compositions and read depths. These features work well on the long contigs, but are not stable for the short ones. Contigs can be linked by sequence overlap (assembly graph) or by the paired-end reads aligned to them (PE graph), where the linked contigs have high chance to be derived from the same clusters. Results We developed METAMVGL, a multi-view graph-based metagenomic contig binning algorithm by integrating both assembly and PE graphs. It could strikingly rescue the short contigs and correct the binning errors from dead ends. METAMVGL learns the two graphs’ weights automatically and predicts the contig labels in a uniform multi-view label propagation framework. In experiments, we observed METAMVGL made use of significantly more high-confidence edges from the combined graph and linked dead ends to the main graph. It also outperformed many state-of-the-art contig binning algorithms, including MaxBin2, MetaBAT2, MyCC, CONCOCT, SolidBin and GraphBin on the metagenomic sequencing data from simulation, two mock communities and Sharon infant fecal samples. Conclusions Our findings demonstrate METAMVGL outstandingly improves the short contig binning and outperforms the other existing contig binning tools on the metagenomic sequencing data from simulation, mock communities and infant fecal samples.

Biosynthetic potential of uncultured Antarctic soil bacteria revealed through long-read metagenomic sequencing

The ISME Journal ◽

10.1038/s41396-021-01052-3 ◽

2021 ◽

Author(s):

Valentin Waschulin ◽

Chiara Borsetto ◽

Robert James ◽

Kevin K. Newsham ◽

Stefano Donadio ◽

...

Keyword(s):

Genome Mining ◽

Gene Clusters ◽

Biosynthetic Gene Cluster ◽

Full Length ◽

Metagenomic Sequencing ◽

Short Read ◽

Short Read Sequencing ◽

Rich Diversity ◽

Long Read ◽

The Rich

AbstractThe growing problem of antibiotic resistance has led to the exploration of uncultured bacteria as potential sources of new antimicrobials. PCR amplicon analyses and short-read sequencing studies of samples from different environments have reported evidence of high biosynthetic gene cluster (BGC) diversity in metagenomes, indicating their potential for producing novel and useful compounds. However, recovering full-length BGC sequences from uncultivated bacteria remains a challenge due to the technological restraints of short-read sequencing, thus making assessment of BGC diversity difficult. Here, long-read sequencing and genome mining were used to recover >1400 mostly full-length BGCs that demonstrate the rich diversity of BGCs from uncultivated lineages present in soil from Mars Oasis, Antarctica. A large number of highly divergent BGCs were not only found in the phyla Acidobacteriota, Verrucomicrobiota and Gemmatimonadota but also in the actinobacterial classes Acidimicrobiia and Thermoleophilia and the gammaproteobacterial order UBA7966. The latter furthermore contained a potential novel family of RiPPs. Our findings underline the biosynthetic potential of underexplored phyla as well as unexplored lineages within seemingly well-studied producer phyla. They also showcase long-read metagenomic sequencing as a promising way to access the untapped genetic reservoir of specialised metabolite gene clusters of the uncultured majority of microbes.

Chloroplast genomes in Populus (Salicaceae): comparisons from an intensively sampled genus reveal dynamic patterns of evolution

Scientific Reports ◽

10.1038/s41598-021-88160-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Jiawei Zhou ◽

Shuo Zhang ◽

Jie Wang ◽

Hongmei Shen ◽

Bin Ai ◽

...

Keyword(s):

Phylogenetic Analyses ◽

Population Level ◽

Gene Content ◽

Sequencing Data ◽

Cellular Functions ◽

Chloroplast Evolution ◽

Chloroplast Genomes ◽

Genome Features ◽

Genome Annotations ◽

Genome Analyses

AbstractThe chloroplast is one of two organelles containing a separate genome that codes for essential and distinct cellular functions such as photosynthesis. Given the importance of chloroplasts in plant metabolism, the genomic architecture and gene content have been strongly conserved through long periods of time and as such are useful molecular tools for evolutionary inferences. At present, complete chloroplast genomes from over 4000 species have been deposited into publicly accessible databases. Despite the large number of complete chloroplast genomes, comprehensive analyses regarding genome architecture and gene content have not been conducted for many lineages with complete species sampling. In this study, we employed the genus Populus to assess how more comprehensively sampled chloroplast genome analyses can be used in understanding chloroplast evolution in a broadly studied lineage of angiosperms. We conducted comparative analyses across Populus in order to elucidate variation in key genome features such as genome size, gene number, gene content, repeat type and number, SSR (Simple Sequence Repeat) abundance, and boundary positioning between the four main units of the genome. We found that some genome annotations were variable across the genus owing in part from errors in assembly or data checking and from this provided corrected annotations. We also employed complete chloroplast genomes for phylogenetic analyses including the dating of divergence times throughout the genus. Lastly, we utilized re-sequencing data to describe the variations of pan-chloroplast genomes at the population level for P. euphratica. The analyses used in this paper provide a blueprint for the types of analyses that can be conducted with publicly available chloroplast genomes as well as methods for building upon existing datasets to improve evolutionary inference.