Descemet Membrane Endothelial Keratoplasty: NGS Metagenomics-Optimized Tissue Preparation and Surgical Technique. Summary of the Doctoral Thesis
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Date
2021
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Rīga Stradiņš University
Abstract
Title: Next-generation sequencing for the detection of microorganisms present in human donor corneal preservation medium. Aim: To detect the presence of microorganisms in the storage media of human donor corneas using next-generation sequencing method. Methods: Seven samples from organ culture (OC) group (Cornea Max, Eurobio, Les Ulis, France) with one control (sterile media without any cornea) and seven samples from hypothermic storage group (Cornea Cold, Eurobio) with one control were used for this study. The corneas were placed in the respective storage media for 14 days before collecting the samples. Storage media (2 mL) from each sample were collected in RNAase-free tubes and shipped for ribosomal RNA sequencing of 16S and 18S. Simultaneously, another 1 mL of media sample was used for conventional diagnostic method (CDM) using Bactec instruments. Results: In both, OC and hypothermic storage and control samples, the most abundant genera were Pseudomonas, Comamonas, Stenotrophomonas, Alcanivorax, Brevundimonas and Nitrobacter. Acidovorax, Acetobacter and Hydrogenophilus were detected mostly in the hypothermic storage group. The most abundant fungal pathogen detected belonged to the genus Malassezia, which was found in both the storage conditions. CDM was negative for microorganisms in all the samples. Conclusion: Metagenomics provides full taxonomic profiling of the detected genomic material of the organisms and thus has the potential to deliver a much wider microbiological diagnostic approach than CDM. The costs and turnaround time need to be reduced, and; the detection of viable organisms would help this technology to be introduced into routine clinical practice.
Description
The Doctoral Thesis was developed at Fondazione Banca degli Occhi del Veneto, Venice, Italy, Rīga Stradiņš University, Riga, Latvia, and at the Department of corneal and external eye diseases, St. Paul's Eye Unit, Royal Liverpool University Hospital, Liverpool, United Kingdom. Defence: at the public session of the Promotion Council of the Clinical Medicine, on 21th December at 12.30 remotely via online platform Zoom.
Keywords
Summary of the Doctoral Thesis, 16S, 18S, NGS, bacteria, cornea, eye bank, fungus, media, microbiology, preservation, storage
Citation
Borroni, D. 2021. Descemet Membrane Endothelial Keratoplasty: NGS Metagenomics-Optimized Tissue Preparation and Surgical Technique: Summary of the Doctoral Thesis: Sub-Sector – Ophthalmology. Rīga: Rīga Stradiņš University. https://doi.org/10.25143/prom-rsu_2021-21_dts