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A Complete Genome Screening Program of Clinical Methicillin-Resistant Staphylococcus aureus Isolates Identifies the Origin and Progression of a Neonatal Intensive Care Unit Outbreak.

Mitchell J Sullivan | Deena R Altman | Kieran I Chacko | Brianne Ciferri | Elizabeth Webster | Theodore R Pak | Gintaras Deikus | Martha Lewis-Sandari | Zenab Khan | Colleen Beckford | Angela Rendo | Flora Samaroo | Robert Sebra | Ramona Karam-Howlin | Tanis Dingle | Camille Hamula | Ali Bashir | Eric Schadt | Gopi Patel | Frances Wallach | Andrew Kasarskis | Kathleen Gibbs | Harm van Bakel
Journal of clinical microbiology | 2019

Whole-genome sequencing (WGS) of Staphylococcus aureus is increasingly used as part of infection prevention practices. In this study, we established a long-read technology-based WGS screening program of all first-episode methicillin-resistant Staphylococcus aureus (MRSA) blood infections at a major urban hospital. A survey of 132 MRSA genomes assembled from long reads enabled detailed characterization of an outbreak lasting several months of a CC5/ST105/USA100 clone among 18 infants in a neonatal intensive care unit (NICU). Available hospital-wide genome surveillance data traced the origins of the outbreak to three patients admitted to adult wards during a 4-month period preceding the NICU outbreak. The pattern of changes among complete outbreak genomes provided full spatiotemporal resolution of its progression, which was characterized by multiple subtransmissions and likely precipitated by equipment sharing between adults and infants. Compared to other hospital strains, the outbreak strain carried distinct mutations and accessory genetic elements that impacted genes with roles in metabolism, resistance, and persistence. This included a DNA recognition domain recombination in the hsdS gene of a type I restriction modification system that altered DNA methylation. Transcriptome sequencing (RNA-Seq) profiling showed that the (epi)genetic changes in the outbreak clone attenuated agr gene expression and upregulated genes involved in stress response and biofilm formation. Overall, our findings demonstrate the utility of long-read sequencing for hospital surveillance and for characterizing accessory genomic elements that may impact MRSA virulence and persistence.

Pubmed ID: 31578260

Associated grants

  • Agency: NIAID NIH HHS, United States
    Id: T32 AI007647
  • Agency: NIAID NIH HHS, United States
    Id: F30 AI122673
  • Agency: NIH HHS, United States
    Id: S10 OD018522
  • Agency: NCATS NIH HHS, United States
    Id: KL2 TR001435
  • Agency: NIAID NIH HHS, United States
    Id: R01 AI119145

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This is a list of tools and resources that we have found mentioned in this publication.


BLASTN (tool)

RRID:SCR_001598

Web application to search nucleotide databases using a nucleotide query. Algorithms: blastn, megablast, discontiguous megablast.

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BLASTX (tool)

RRID:SCR_001653

Web application to search protein databases using a translated nucleotide query. Translated BLAST services are useful when trying to find homologous proteins to a nucleotide coding region. Blastx compares translational products of the nucleotide query sequence to a protein database. Because blastx translates the query sequence in all six reading frames and provides combined significance statistics for hits to different frames, it is particularly useful when the reading frame of the query sequence is unknown or it contains errors that may lead to frame shifts or other coding errors. Thus blastx is often the first analysis performed with a newly determined nucleotide sequence and is used extensively in analyzing EST sequences. This search is more sensitive than nucleotide blast since the comparison is performed at the protein level.

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RAxML (tool)

RRID:SCR_006086

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Bioconductor (tool)

RRID:SCR_006442

Software repository for R packages related to analysis and comprehension of high throughput genomic data. Uses separate set of commands for installation of packages. Software project based on R programming language that provides tools for analysis and comprehension of high throughput genomic data.

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FreeBayes (tool)

RRID:SCR_010761

A Bayesian genetic variant detector designed to find small polymorphisms, specifically SNPs, indels, MNPs, and complex events smaller than the length of a short-read sequencing alignment.

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edgeR (tool)

RRID:SCR_012802

Bioconductor software package for Empirical analysis of Digital Gene Expression data in R. Used for differential expression analysis of RNA-seq and digital gene expression data with biological replication.

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PubMLST (tool)

RRID:SCR_012955

Database for molecular typing and microbial genome diversity.

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Easyfig (tool)

RRID:SCR_013169

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Prokka (tool)

RRID:SCR_014732

THIS RESOURCE IS NO LONGER IN SERVICE. Documented on February 28,2023. Software tool for the rapid annotation of prokaryotic genomes. It produces GFF3, GBK and SQN files that are ready for editing in Sequin and ultimately submitted to Genbank/DDJB/ENA. A typical 4 Mbp genome can be fully annotated in less than 10 minutes on a quad-core computer, and scales well to 32 core SMP systems.

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Circlator (tool)

RRID:SCR_016058

Software that automates assembly circularization and produces accurate linear representations of circular sequences. It is used for assembling of DNA sequence data of complete bacterial and small eukaryotic genomes.

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NetworkX (tool)

RRID:SCR_016864

Software Python package for the creation, manipulation, and study of the structure, dynamics, and functions of complex networks.

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BWA-MEM2 (tool)

RRID:SCR_022192

Software tool for sequence mapping.The next version of BWA-MEM. Used for aligning sequencing reads against large reference genome.

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IGB (tool)

RRID:SCR_011792

An easy-to-use, highly customizable genome browser you can use to visualize and explore genomic data and annotations, including RNA-Seq, ChIP-Seq, tiling array data, and more.

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