Department of Ophthalmology, Faculty of Medicine and Health, Örebro University, SE 701 82, Örebro, Sweden.
Department of Bacteria, Parasites and Fungi, Statens Serum Institut, Copenhagen, Denmark.
Graefes Arch Clin Exp Ophthalmol. 2024 May;262(5):1579-1589. doi: 10.1007/s00417-023-06310-y. Epub 2023 Nov 23.
To describe the bacterial findings by a targeted sequencing approach from corneal samples of patients with microbial keratitis and factors influencing culture outcome of indirectly inoculated corneal specimen.
Prospective inclusion of patients fulfilling predefined criteria of microbial keratitis. Samples from the corneal lesion were collected and dispensed in liquid transport medium, from which both culture and targeted amplification and sequencing of the V3-V4 region of the 16S rRNA gene were carried out. Additional standard corneal culture from the corneal lesions was also performed. Factors influencing culture outcome of indirectly inoculated corneal samples were identified by a multivariate regression model incorporating quantitative data from sequencing.
Among the 94 included patients with microbial keratitis, contact lens wear (n = 69; 73%) was the most common risk factor. Contact lens wearers displayed significant differences in the bacterial community composition of the corneal lesion compared to no lens wearers, with higher abundance of Staphylococcus spp., Corynebacterium spp., and Stenotrophomonas maltophilia. Targeted sequencing detected a potential corneal pathogen in the highest proportional abundance among 9 of the 24 (38%) culture-negative patients with microbial keratitis. Age, bacterial density in the sample, and prior antibiotic treatment significantly influenced culture outcome of indirectly inoculated corneal samples.
Targeted sequencing may provide insights on pathogens in both culture negative episodes of microbial keratitis and among subgroups of patients with microbial keratitis as well as factors influencing culture outcome of indirectly inoculated corneal samples.
通过靶向测序方法描述微生物角膜炎患者的角膜样本中的细菌发现，并探讨影响间接接种角膜标本培养结果的因素。
前瞻性纳入符合微生物角膜炎预设标准的患者。采集角膜病变部位的样本并置于液体运输培养基中，对其进行培养，并对 16S rRNA 基因的 V3-V4 区进行靶向扩增和测序。此外，还对角膜病变处进行了常规的角膜培养。通过多元回归模型，结合测序的定量数据，确定影响间接接种角膜样本培养结果的因素。
在 94 例患有微生物角膜炎的患者中，最常见的危险因素是隐形眼镜佩戴（n=69；73%）。与不佩戴隐形眼镜的患者相比，隐形眼镜佩戴者的角膜病变部位的细菌群落组成存在显著差异，其中金黄色葡萄球菌、棒状杆菌属和嗜麦芽窄食单胞菌的丰度更高。在 24 例（38%）培养阴性的微生物角膜炎患者中，有 9 例患者的靶向测序检测到了最高比例的潜在角膜病原体。年龄、样本中的细菌密度以及既往抗生素治疗显著影响了间接接种角膜样本的培养结果。
靶向测序可能为培养阴性的微生物角膜炎发作以及微生物角膜炎患者亚组中病原体的鉴定以及影响间接接种角膜样本培养结果的因素提供新的见解。
