Complete Genome Sequence of Staphylococcus aureus CI/BAC/25/13/W, Isolated from Contaminated Platelet Concentrates in England

Paredes, Carina; Ighem, Sylvia; Flint, Annika; Weedmark, Kelly; McDonald, Carl; Bearne, Jennifer; Ramirez‐Arcos, Sandra; Pagotto, Franco

doi:10.1128/mra.00840-21

Cited by 2 publications

(5 citation statements)

References 10 publications

(10 reference statements)

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“…The third English strain (CI/BAC/25/13/W) and the Canadian isolate (CBS2016‐05) were missed during BACT/ALERT PC screening (i.e., false negative screening). While the PC unit contaminated with S. aureus CI/BAC/25/13/W was identified prior to transfusion by visual inspection due to clot formation (‘near‐miss’) [12], the PC unit carrying CBS2016‐05 was transfused causing a septic transfusion reaction [14].…”

Section: Methodsmentioning

confidence: 99%

“…Four S. aureus strains listed in Table 1 were used in this study, including one isolated in Canada [9] and three isolated in England [10][11][12]. Two of the English isolates (PS/BAC/169/17/W and PS/BAC/317/16/W) were detected during routine PC screening using the BACT/ALERT culture system at the National Health Service Blood and Transplant (NHSBT) (i.e., confirmed positives).…”

Section: S Aureus Isolatesmentioning

confidence: 99%

“…The genomes of all four S. aureus isolates were next-generation sequenced. Illumina paired end DNA libraries of the isolates were sequenced using the Miseq platform, assembled, annotated at the Health Canada genomics facility, and published at the NCBI database [9][10][11][12]. The presence of SE genes in the next-generation sequences of all studied strains was further confirmed by Sanger sequencing.…”

Section: Staphylococcal Exotoxin Gene Identificationmentioning

confidence: 99%

“…Gram-positive and gram-negative bacteria have been isolated from PCs with Staphylococcus aureus ranking as a major cause of transfusion related sepsis and fatalities [2,[9][10][11][12][13][14]. This species can evade detection during routine PC screening with automated culture systems due to slow growth and ability to form biofilms in PCs [13,14].…”

Section: Introductionmentioning

confidence: 99%

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Proof of concept for detection of staphylococcal enterotoxins in platelet concentrates as a novel safety mitigation strategy

et al. 2023

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Background and Objectives Staphylococcus aureus is a predominant contaminant of platelet concentrates (PCs) that can evade detection during screening with culture methods. Importantly, S. aureus produces staphylococcal enterotoxins (SEs) during PC storage, which are linked to slow growth and enhanced biofilm formation. This study investigated timing of SE production during PC storage and feasibility of SE detection as a PC safety strategy. Materials and Methods Genomic and transcriptomic data of transfusion‐relevant S. aureus PS/BAC/169/17/W, PS/BAC/317/16/W, CI/BAC/25/13/W and CBS2016‐05 were used to determine the presence and differential expression of exotoxin genes in PCs. Trypticase soy broth (TSB) and PCs were inoculated with 1.0E+06 cfu/mL of S. aureus PS/BAC/169/17/W and CBS2016‐05. Expression of SEs at different growth phases was confirmed with Western blotting. PCs were inoculated with 30 cfu/unit of the same strains, and SE detection during PC storage was optimized with a sandwich dot‐ELISA assay. Results S. aureus genomes contain multiple exotoxin genes including those encoding for SEs. Transcriptome data revealed significant upregulation (0.5–6.7‐fold, p < 0.05) of SE genes in PCs versus TSB. Western blots demonstrated SE production at all growth phases. Notably, dot‐ELISA detected clinically relevant concentrations of SEs (~0.2 μg/mL) at 32 h of PC storage when S. aureus PS/BAC/169/17/W and CBS2016‐05 counts were 1.8E+04 and 1.4E+04 cfu/mL, respectively. Conclusion Genomic analyses revealed that staphylococcal exotoxins are widely distributed and highly conserved among transfusion‐relevant S. aureus isolates. Furthermore, SEs are significantly upregulated in PCs and detected at 30 h of PC storage. Therefore, bacterial toxin detection could supplement mitigation strategies to enhance PC safety.

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Section: Methodsmentioning

confidence: 99%

Section: S Aureus Isolatesmentioning

confidence: 99%

Section: Staphylococcal Exotoxin Gene Identificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Proof of concept for detection of staphylococcal enterotoxins in platelet concentrates as a novel safety mitigation strategy

et al. 2023

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“…The S. aureus strains used in this study were isolated from contaminated PCs in Canada and England as recently described [ 20 , 21 , 22 , 23 ]. Two of the isolates, CBS2016-05 and PS/BAC/169/17/W, are strong biofilm producers, while the other two strains, CI/BAC/25/13/W and PS/BAC/317/16/W, form weak biofilms [ 24 ].…”

Section: Methodsmentioning

confidence: 99%

Staphylococcal Enterotoxins Enhance Biofilm Formation by Staphylococcus aureus in Platelet Concentrates

Ighem

Ramirez‐Arcos

2022

Microorganisms

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Biofilm formation and slow growth by Staphylococcus aureus in platelet concentrates (PCs) cause missed detection of this bacterium during routine PC screening with automated culture systems. This heightens the chances of false-negative screening transfusions and pre-disposes transfusion patients to an elevated risk of sepsis due to secretion of staphylococcal enterotoxins (SEs) in PCs. A hybrid approach of comparative RNAseq analyses and CRISPR mutagenesis of SE genes was employed to investigate the effect of SEs in S. aureus growth and biofilm formation in PCs. RNAseq data showed no differential expression for key biofilm genes, whereas SE genes were upregulated (>0.5- to 3.6-fold change) in PCs compared to trypticase soy broth (TSB). Remarkably, growth and biofilm formation assays revealed increased growth for the S. aureus SE mutants, while their ability to form biofilms was significantly impaired (−6.8- to −2.4-fold change) in comparison to the wild type strain, in both PCs and TSB. Through the well-established superantigen mechanism of SEs, we propose three roles for SEs during biofilm development in PCs: (1) provide a scaffold for biofilm matrix, (2) mediate cell-to-cell aggregation, and (3) guarantee biofilm survival. Furthermore, SE contribution to both growth and biofilm development seems to be centrally regulated by agr via quorum sensing and by saeSR and sigB. This study reveals new roles for SEs, which enforce their relevance in ensuring PC safety for transfusion patients. It further deciphers the underlying reasons for failed S. aureus detection in PCs during screening with automated culture systems.

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Complete Genome Sequence of Staphylococcus aureus CI/BAC/25/13/W, Isolated from Contaminated Platelet Concentrates in England

Cited by 2 publications

References 10 publications

Proof of concept for detection of staphylococcal enterotoxins in platelet concentrates as a novel safety mitigation strategy

Proof of concept for detection of staphylococcal enterotoxins in platelet concentrates as a novel safety mitigation strategy

Staphylococcal Enterotoxins Enhance Biofilm Formation by Staphylococcus aureus in Platelet Concentrates

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