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. 2017 Mar 30;5(13):e00098-17. doi: 10.1128/genomeA.00098-17

Draft Genome Sequences of 14 Staphylococcus aureus Sequence Type 5 Isolates from California, USA

Samantha J Hau a, Darrell O Bayles b, David P Alt b, Tracy L Nicholson b,
PMCID: PMC5374240  PMID: 28360166

ABSTRACT

Staphylococcus aureus is part of the human epithelial microbiota; however, it is also a pathogen. The acquisition of mobile genetic elements plays a role in the virulence of S. aureus isolates and contributes to treatment failures. This report details the draft genome sequences of 14 clinical S. aureus isolates.

GENOME ANNOUNCEMENT

Staphylococcus aureus is part of the microbiota found on the skin of humans and other animals. It is also a known pathogen that causes mild to severe infections, as well as toxin-mediated diseases (1). Methicillin-resistant S. aureus (MRSA) is a growing concern in the health industry, and MRSA isolates are categorized based on each isolate’s source as either hospital-acquired MRSA (HA-MRSA), community-acquired MRSA (CA-MRSA), or livestock-associated MRSA (LA-MRSA). Multilocus sequence typing is employed to determine an isolate’s sequence type (ST), which indicates the genetic background and denotes the characteristics of the isolate common to the group. The ST5 lineage is globally distributed and well known for acquisition of virulence factors and antibiotic resistance genes contained on mobile genetic elements (2).

We sequenced 14 ST5 isolates from patients with clinical disease caused by MRSA. These isolates were obtained from the University of California, Irvine (UCI3, UCI9, UCI11, UCI19, UCI21, UCI24, UCI27, UCI43, UCI45, UCI46, UCI48, UCI52, UCI56, and UCI64) (3). The patients had no known livestock exposure; however, because full patient histories were not provided, it could not be determined whether the isolates were HA- or CA-MRSA. Isolates were grown in Trypticase soy broth (BD Biosciences, Sparks, MD), and total genomic DNA was extracted using the High Pure PCR template preparation kit (Roche Applied Science, Indianapolis, IN).

Draft genome sequences were generated using an Illumina MiSeq instrument. Nextera XT DNA sample preparation and index kits (Illumina, San Diego, CA) were used to generate indexed libraries. Libraries were pooled and sequenced using the MiSeq version 2 500-cycle reagent kit, yielding 2 × 250-bp paired-end reads (Illumina).

Draft genome assemblies were generated using MIRA version 4.0.2 (http://mira-assembler.sourceforge.net/docs/DefinitiveGuideToMIRA.html), resulting in the average coverages indicated here for each isolate: UCI3, 56×; UCI9, 82×; UCI11, 72×; UCI19, 47×; UCI21, 66×; UCI24, 62×; UCI27, 40×; UCI43, 46×; UCI45, 53×; UCI46, 73×; UCI48, 46×; UCI52, 33×; UCI56, 36×; and UCI64, 52×. Following assembly, only contigs >1,500 bp in length having a coverage of >66% of the average genome coverage were retained. Additionally, when the assembly tool indicated that a contig was part of a potentially repetitive element, the contig was required to be >2,000 bp for inclusion in the assembly.

Accession number(s).

The draft genome sequences obtained for these isolates were entered into DDBJ/ENA/GenBank with accession numbers as follows: UCI3, LKYU00000000; UCI9, LKZA00000000; UCI11, LKZC00000000; UCI19, LKZK00000000; UCI21, LKZM00000000; UCI24, LKZP00000000; UCI27, LKZS00000000; UCI43, LLAI00000000; UCI45, LLAK00000000; UCI46, LLAL00000000; UCI48, LLAN00000000; UCI52, LLAR00000000; UCI56, LLAV00000000; and UCI64, LLBD00000000.

ACKNOWLEDGMENTS

The research herein was funded by the United States Department of Agriculture, Agricultural Research Service, and by the National Pork Board. S.J.H. is supported by the Iowa State University, Veterinary Diagnostic Laboratory. The funding sources had no role in the study design, data collection or analysis, decision to publish, or manuscript preparation.

The mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer.

Footnotes

Citation Hau SJ, Bayles DO, Alt DP, Nicholson TL. 2017. Draft genome sequences of 14 Staphylococcus aureus sequence type 5 isolates from California, USA. Genome Announc 5:e00098-17. https://doi.org/10.1128/genomeA.00098-17.

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