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

Author:

Chi Sylvia Ighem12,Yousuf Basit12,Paredes Carina12,Bearne Jennifer3,McDonald Carl3ORCID,Ramirez‐Arcos Sandra12ORCID

Affiliation:

1. Medical Affairs and Innovation Canadian Blood Services Ottawa Ontario Canada

2. Department of Biochemistry, Microbiology and Immunology University of Ottawa Ottawa Ontario Canada

3. National Health Service Blood and Transplant London UK

Abstract

AbstractBackground and ObjectivesStaphylococcus 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 MethodsGenomic 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.ResultsS. 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.ConclusionGenomic 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.

Funder

Canadian Blood Services

Health Canada

Publisher

Wiley

Subject

Hematology,General Medicine

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