Centre for Innovation, Canadian Blood Services, Ottawa, Ontario, Canada.
Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario, Canada.
Transfusion. 2021 Jul;61(7):2146-2158. doi: 10.1111/trf.16418. Epub 2021 May 17.
Staphylococcus epidermidis forms surface-attached aggregates (biofilms) when grown in platelet concentrates (PCs). Comparative transcriptome analyses were undertaken to investigate differential gene expression of S. epidermidis biofilms grown in PCs.
Two S. epidermidis strains isolated from human skin (AZ22 and AZ39) and one strain isolated from contaminated PCs (ST02) were grown in glucose-supplemented Trypticase Soy Broth (TSBg) and PCs. RNA was extracted and sequenced using Illumina HiSeq. Differential expression analysis was done using DESeq, and significantly differentially expressed genes (DEGs) were selected. DEGs were subjected to Kyoto encyclopedia of genes and genomes and Gene Ontology analyses. Differential gene expression was validated with quantitative reverse transcription-PCR.
A total of 436, 442, and 384 genes were expressed in AZ22, AZ39, and ST02, respectively. DEG analysis showed that 170, 172, and 117 genes were upregulated in PCs in comparison to TSBg, whereas 120, 135, and 89 genes were downregulated (p < .05) in mature biofilms of AZ22, AZ39, and ST02, respectively. Twenty-seven DEGs were shared by all three strains. While 76 DEGs were shared by AZ22 and AZ39, only 34 and 21 DEGs were common between ST02, and AZ22 and AZ39, respectively. Significant transcriptional expression changes were observed in genes involved in platelet-bacteria interaction, biofilm formation, production of virulence factors, and resistance to antimicrobial peptides and antibiotics.
Differential gene expression in S. epidermidis is triggered by the stressful PC storage environment. Upregulation of virulence and antimicrobial resistance genes could have clinical implications for transfusion patients.
表皮葡萄球菌在血小板浓缩物(PCs)中生长时会形成附着在表面的聚集物(生物膜)。进行比较转录组分析,以研究在 PCs 中生长的表皮葡萄球菌生物膜的差异基因表达。
从人类皮肤中分离出的两株表皮葡萄球菌(AZ22 和 AZ39)和一株从污染的 PCs 中分离出的菌株(ST02)在葡萄糖补充的胰蛋白酶大豆肉汤(TSBg)和 PCs 中生长。使用 Illumina HiSeq 提取和测序 RNA。使用 DESeq 进行差异表达分析,并选择显著差异表达基因(DEGs)。对 DEGs 进行京都基因与基因组百科全书和基因本体论分析。用定量逆转录-PCR 验证差异基因表达。
AZ22、AZ39 和 ST02 分别表达了 436、442 和 384 个基因。DEG 分析表明,与 TSBg 相比,170、172 和 117 个基因在 PCs 中上调,而 AZ22、AZ39 和 ST02 成熟生物膜中分别有 120、135 和 89 个基因下调(p<.05)。三个菌株共有的 27 个 DEG。AZ22 和 AZ39 共有 76 个 DEG,而 ST02 与 AZ22 和 AZ39 分别有 34 和 21 个 DEG 共有。参与血小板-细菌相互作用、生物膜形成、毒力因子产生以及对抗生素和抗生素耐药性的基因的转录表达发生了显著变化。
表皮葡萄球菌的差异基因表达是由应激性 PC 储存环境引发的。毒力和抗药性基因的上调可能对输血患者具有临床意义。