Department of Ophthalmology, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
Department of Laboratory Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
Int J Mol Sci. 2020 Nov 5;21(21):8299. doi: 10.3390/ijms21218299.
A sound ocular surface microbiota has been recognized as a part of ocular surface health following a growing body of evidence from next-generation sequencing technique and metagenomic analysis. However, even from the perspective of contemporary precision medicine, it is difficult to directly apply these new technologies to clinical practice. Therefore, we proposed a model based on dot hybridization assay (DHA) to bridge conventional culture with a metagenomic approach in investigating and monitoring ocular surface microbiota. Endophthalmitis, mostly caused by bacterial infection, is the most severe complication of many intraocular surgeries, such as cataract surgery. Hazardous microorganisms hiding and proliferating in the ocular surface microbiota not only increase the risk of endophthalmitis but also jeopardize the effectiveness of the preoperative aseptic procedure and postoperative topical antibiotics. The DHA model enables the simultaneous assessment of bacterial bioburden, detection of target pathogens and microorganisms, and surveillance of methicillin/oxacillin resistance gene in the ocular surface microbiota. This assay revealed heavier bacterial bioburden in men, compatible with a higher risk of endophthalmitis in male patients who underwent cataract surgery. No occurrence of endophthalmitis for these patients was compatible with non-hazardous microorganisms identified by specific dots for target pathogens. Moreover, the mecA dot detected oxacillin-resistant strains, of which culture failed to isolate. Therefore, the DHA model could provide an alternative genomic approach to investigate and monitor ocular surface microorganisms in clinical practice nowadays.
越来越多的下一代测序技术和宏基因组分析证据表明，健康的眼表面需要有良好的眼表面微生物群。然而，即使从当代精准医学的角度来看，也很难将这些新技术直接应用于临床实践。因此，我们提出了一种基于点杂交检测（DHA）的模型，该模型将常规培养与宏基因组方法相结合，用于调查和监测眼表面微生物群。内源性眼炎主要由细菌感染引起，是许多眼内手术（如白内障手术）最严重的并发症。隐藏和增殖在眼表面微生物群中的有害微生物不仅增加了内源性眼炎的风险，而且危及术前无菌程序和术后局部抗生素的有效性。DHA 模型能够同时评估细菌生物负荷、检测目标病原体和微生物，并监测眼表面微生物群中的耐甲氧西林/苯唑西林基因。该检测结果显示，男性的细菌生物负荷更高，与男性白内障手术患者内源性眼炎风险更高的情况相符。这些患者没有发生内源性眼炎，这与针对目标病原体的特定斑点识别的非危险微生物相符。此外，mecA 斑点检测到耐苯唑西林的菌株，但培养未能分离出这些菌株。因此，DHA 模型可以为当今临床实践中调查和监测眼表面微生物提供一种替代的基因组方法。
