scholarly journals Near-Complete Genome Sequence of a Human Norovirus GII.1[Pg] Strain Associated with Acute Gastroenteritis, Determined Using Long-Read Sequencing

2021 ◽  
Vol 10 (28) ◽  
Author(s):  
Zhihui Yang ◽  
Mark Mammel ◽  
Samantha Q. Wales
Keyword(s):  
Foodborne Pathogens ◽  
High Throughput Sequencing ◽  
Read Length ◽  
Human Norovirus ◽  
Short Read ◽  
Fast Detection ◽  
Short Read Sequencing ◽  
Detection And Identification ◽  
Long Read ◽  
Norovirus Gii

High-throughput sequencing is one of the approaches used for the detection of foodborne pathogens such as noroviruses. Long-read sequencing has advantages over short-read sequencing in speed, read length, and lower fragmentation bias, which makes it a potential powerful tool for the fast detection and identification of viruses.

scholarly journals rRNA Analysis Based on Long-Read High-Throughput Sequencing Reveals a More Accurate Diagnostic for the Bacterial Infection of Ascites

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Xiaoling Yu ◽  
Wenqian Jiang ◽  
Xinhui Huang ◽  
Jun Lin ◽  
Hanhui Ye ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
High Throughput Sequencing ◽  
Second Generation ◽  
Subsequent Treatment ◽  
Microbial Culture ◽  
Third Generation ◽  
Short Read ◽  
Short Read Sequencing ◽  
Long Read ◽  
Generation Sequencing

Traditional pathogenic diagnosis presents defects such as a low positivity rate, inability to identify uncultured microorganisms, and time-consuming nature. Clinical metagenomics next-generation sequencing can be used to detect any pathogen, compensating for the shortcomings of traditional pathogenic diagnosis. We report third-generation long-read sequencing results and second-generation short-read sequencing results for ascitic fluid from a patient with liver ascites and compared the two types of sequencing results with the results of traditional clinical microbial culture. The distribution of pathogenic microbial species revealed by the two types of sequencing results was quite different, and the third-generation sequencing results were consistent with the results of traditional microbial culture, which can effectively guide subsequent treatment. Short reads, the lack of amplification, and enrichment to amplify signals from trace pathogens, and host background noise may be the reasons for the high error in the second-generation short-read sequencing results. Therefore, we propose that long-read-based rRNA analysis technology is superior to the short-read shotgun-based metagenomics method in the identification of pathogenic bacteria.

scholarly journals rRNA analysis based on long-read high-throughput sequencing reveals a more accurate diagnostic for the bacterial infection of ascites

2020 ◽  
Author(s):  
Xiaoling Yu ◽  
Wenqian Jiang ◽  
Xinhui Huang ◽  
Jun Lin ◽  
Hanhui Ye ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
High Throughput Sequencing ◽  
Second Generation ◽  
Subsequent Treatment ◽  
Microbial Culture ◽  
Third Generation ◽  
Short Read ◽  
Short Read Sequencing ◽  
Long Read ◽  
Generation Sequencing

Traditional pathogenic diagnosis presents defects such as a low positivity rate, inability to identify uncultured microorganisms, and time-consuming nature. Clinical metagenomics next-generation sequencing can be used to detect any pathogen, compensating for the shortcomings of traditional pathogenic diagnosis. We report third-generation long-read sequencing results and second-generation short-read sequencing results for ascitic fluid from a patient with liver ascites and compared the two types of sequencing results with the results of traditional clinical microbial culture. The distribution of pathogenic microbial species revealed by the two types of sequencing results was quite different, and the third-generation sequencing results were consistent with the results of traditional microbial culture, which can effectively guide subsequent treatment. Short reads, the lack of amplification and enrichment to amplify signals from trace pathogens, and host background noise may be the reasons for high error in the second-generation short-read sequencing results. Therefore, we propose that long-read-based rRNA analysis technology is superior to the short-read shotgun-based metagenomics method in the identification of pathogenic bacteria.

scholarly journals Closed Genome Sequences of 28 Foodborne Pathogens from the CFSAN Verification Set, Determined by a Combination of Long and Short Reads

2020 ◽  
Vol 9 (18) ◽  
Author(s):  
Narjol Gonzalez-Escalona ◽  
George John Kastanis ◽  
Ruth Timme ◽  
Dwayne Roberson ◽  
Maria Balkey ◽  
...  
Keyword(s):  
Foodborne Pathogens ◽  
Bacterial Strains ◽  
Genome Sequences ◽  
Short Read ◽  
Short Reads ◽  
Short Read Sequencing ◽  
Long Read

Foodborne pathogens have been implicated in illnesses worldwide. Here, we report the complete closed genome sequences of 28 bacterial strains belonging to 18 different species. These genomes belong to known foodborne pathogens. The genomes were closed by a combination of long-read and short-read sequencing.

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

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.

scholarly journals Rapid Mycobacterium tuberculosis spoligotyping from uncorrected long reads using Galru

2020 ◽  
Author(s):  
Andrew J. Page ◽  
Nabil-Fareed Alikhan ◽  
Michael Strinden ◽  
Thanh Le Viet ◽  
Timofey Skvortsov
Keyword(s):  
Mycobacterium Tuberculosis ◽  
State Of The Art ◽  
Sequence Data ◽  
Human Pathogen ◽  
Sequencing Data ◽  
Short Read ◽  
Short Read Sequencing ◽  
Long Reads ◽  
Long Read

AbstractSpoligotyping of Mycobacterium tuberculosis provides a subspecies classification of this major human pathogen. Spoligotypes can be predicted from short read genome sequencing data; however, no methods exist for long read sequence data such as from Nanopore or PacBio. We present a novel software package Galru, which can rapidly detect the spoligotype of a Mycobacterium tuberculosis sample from as little as a single uncorrected long read. It allows for near real-time spoligotyping from long read data as it is being sequenced, giving rapid sample typing. We compare it to the existing state of the art software and find it performs identically to the results obtained from short read sequencing data. Galru is freely available from https://github.com/quadram-institute-bioscience/galru under the GPLv3 open source licence.

scholarly journals Closed Genome Sequence of Salmonella enterica Serovar Richmond Strain CFSAN000191, Obtained with Nanopore Sequencing

2018 ◽  
Vol 7 (23) ◽  
Author(s):  
Narjol González-Escalona ◽  
Kuan Yao ◽  
Maria Hoffmann
Keyword(s):  
Genome Sequence ◽  
Salmonella Enterica ◽  
Nanopore Sequencing ◽  
Short Read ◽  
Content Type ◽  
Short Read Sequencing ◽  
Long Read

Here we report the genome sequence of Salmonella enterica serovar Richmond strain CFSAN000191, isolated from tilapia from Thailand in 2005. The genome was determined by a combination of long-read and short-read sequencing.

scholarly journals Closed Genome Sequences of Three Salmonella enterica Strains Belonging to Serovars Saintpaul, Weltevreden, and Thompson, Isolated from Mexico

2019 ◽  
Vol 8 (28) ◽  
Author(s):  
Narjol Gonzalez-Escalona ◽  
J. R. Aguirre-Sánchez ◽  
J. R. Ibarra-Rodríguez ◽  
C. Chaidez-Quiroz ◽  
Jaime Martinez-Urtaza
Keyword(s):  
River Water ◽  
Salmonella Enterica ◽  
Genome Sequences ◽  
Short Read ◽  
Content Type ◽  
Short Read Sequencing ◽  
Long Read ◽  
Sequence Types

Here, we report the genome sequences of three Salmonella enterica strains belonging to serovars Weltevreden (CFSAN047349), Saintpaul (CFSAN047351), and Thompson (CFSAN047352), isolated from river water in Sinaloa, Mexico. The genomes were closed by a combination of long-read and short-read sequencing. The strain sequence types (STs) are ST365, ST50, and ST26, respectively.

scholarly journals Complete Genome Sequence of Rubrobacter xylanophilus Strain AA3-22, Isolated from Arima Onsen in Japan

2019 ◽  
Vol 8 (34) ◽  
Author(s):  
Natsuki Tomariguchi ◽  
Kentaro Miyazaki
Keyword(s):  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Hot Spring ◽  
Sequencing Data ◽  
Short Read ◽  
Content Type ◽  
Short Read Sequencing ◽  
Oxford Nanopore ◽  
Long Read

Rubrobacter xylanophilus strain AA3-22, belonging to the phylum Actinobacteria, was isolated from nonvolcanic Arima Onsen (hot spring) in Japan. Here, we report the complete genome sequence of this organism, which was obtained by combining Oxford Nanopore long-read and Illumina short-read sequencing data.

scholarly journals Towards a Comprehensive Variation Benchmark for Challenging Medically-Relevant Autosomal Genes

2021 ◽  
Author(s):  
Justin Wagner ◽  
Nathan D Olson ◽  
Lindsay Harris ◽  
Jennifer McDaniel ◽  
Haoyu Cheng ◽  
...  
Keyword(s):  
Genome Sequencing ◽  
Method Development ◽  
Clinical Applications ◽  
Accurate Analysis ◽  
Short Read ◽  
Short Read Sequencing ◽  
Repetitive Nature ◽  
Long Read ◽  
Comprehensive Characterization

The repetitive nature and complexity of multiple medically important genes make them intractable to accurate analysis, despite the maturity of short-read sequencing, resulting in a gap in clinical applications of genome sequencing. The Genome in a Bottle Consortium has provided benchmark variant sets, but these excluded some medically relevant genes due to their repetitiveness or polymorphic complexity. In this study, we characterize 273 of these 395 challenging autosomal genes that have multiple implications for medical sequencing. This extended, curated benchmark reports over 17,000 SNVs, 3,600 INDELs, and 200 SVs each for GRCh37 and GRCh38. We show that false duplications in either GRCh37 or GRCh38 result in reference-specific, missed variants for short- and long-read technologies in medically important genes including CBS, CRYAA, and KCNE1. Our proposed solution improves variant recall in these genes from 8% to 100%. This benchmark will significantly improve the comprehensive characterization of these medically relevant genes and guide new method development.

scholarly journals LinkedSV for detection of mosaic structural variants from linked-read exome and genome sequencing data

2018 ◽  
Author(s):  
Li Fang ◽  
Charlly Kao ◽  
Michael V Gonzalez ◽  
Fernanda A Mafra ◽  
Renata Pellegrino da Silva ◽  
...  
Keyword(s):  
Exome Sequencing ◽  
Read Depth ◽  
Structural Variants ◽  
Sequencing Data ◽  
High Coverage ◽  
Short Read ◽  
Short Read Sequencing ◽  
Sequencing Studies ◽  
Long Read ◽  
Local Assembly

AbstractLinked-read sequencing provides long-range information on short-read sequencing data by barcoding reads originating from the same DNA molecule, and can improve the detection and breakpoint identification for structural variants (SVs). We present LinkedSV for SV detection on linked-read sequencing data. LinkedSV considers barcode overlapping and enriched fragment endpoints as signals to detect large SVs, while it leverages read depth, paired-end signals and local assembly to detect small SVs. Benchmarking studies demonstrates that LinkedSV outperforms existing tools, especially on exome data and on somatic SVs with low variant allele frequencies. We demonstrate clinical cases where LinkedSV identifies disease causal SVs from linked-read exome sequencing data missed by conventional exome sequencing, and show examples where LinkedSV identifies SVs missed by high-coverage long-read sequencing. In summary, LinkedSV can detect SVs missed by conventional short-read and long-read sequencing approaches, and may resolve negative cases from clinical genome/exome sequencing studies.

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