Abstract
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 mecA 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.
Highlights
The microbiome has been linked to cancer, obesity, asthma, rheumatoid arthritis, and diabetes, illustrating the significance of gut microbiota in health and illness [1,2,3,4]
Observing the very low signal intensities of the dots for all target microorganisms, the result was compatible with low risk of infection for these patients, who had no occurrence of endophthalmitis after cataract surgery
The Dot Hybridization Assay for Assessing COSM. In this specially-designed dot hybridization assay (DHA) (Figure 1, Table 1) for assessing COSM, there were one bacteria universal dot (BPx; designed from conserved sequences at the 3 end of the 16S rRNA gene) for estimating overall bacterial bioburden, seven bacteria-specific dots for detecting bacteria of putative core ocular surface microbiota, and one dot for surveilling antibiotic resistance of ocular surface bacteria
Summary
The microbiome has been linked to cancer, obesity, asthma, rheumatoid arthritis, and diabetes, illustrating the significance of gut microbiota in health and illness [1,2,3,4]. Unlike any other body sites, the heathy conjunctiva, lid margins, and tears have remarkably fewer microbial species than other mucosa sites, such as oral mucosa surface [7]. Considering the limitations from the above modalities in understanding microbiota on personal ocular surface, the DNA dot hybridization assay (DHA) may provide a more rational way by detecting target microorganisms [18,19], quantifying microbial burden [20,21], and surveilling antibiotic resistance [22] if the core ocular surface microbiota (COSM) is defined. We proposed a postulated COSM, in which target microorganisms are more frequently recovered or recognized as pathogens or flora from the ocular surface by culture and Sanger sequencing methods. This study selected patients who underwent cataract surgeries in order to explore the microorganism profile in the COSM using the DHA model
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