ESCRS - PP17.10 - Shotgun Metagenomic Sequencing In Culture Negative Microbial Keratitis.

Shotgun Metagenomic Sequencing In Culture Negative Microbial Keratitis.

Published 2022 - 40th Congress of the ESCRS

Reference: PP17.10 | Type: Free paper | DOI: 10.82333/k8p3-2v12

Authors: Davide Borroni* 1 , Chiara Bonzano 2 , Carlos Rocha de Lossada 3 , Maria Garcia Lorente 4 , Fernando Godin 5 , Jose'-Maria Sánchez-González 6 , Marina Rodríguez-Calvo-de-Mora 4

1Ophthalmology,VISIA CLINIC,MILAN,Italy, 2Ophthalmology,DiNOGMI, University of Genoa and IRCCS San Martino Polyclinic Hospital,Genoa,Italy, 3Department of Ophthalmology ,University Hospital Virgen,Granada,Spain, 4Department of Ophthalmology,Hospital Regional Universitario Málaga,Malaga,Spain, 5Department of Ophthalmology,Universidad el Bosque ,Bogota',Colombia, 6Department of Physics of Condensed Matter,University of Seville,Seville,Spain

Purpose

To evaluate the microbiota of culture negative Corneal Impression Membrane (CIM) microbial keratitis samples with the use of shotgun metagenomics analysis.

Setting

Department of Doctoral Studies, Riga Stradins University, Riga, Latvia;

DiNOGMI, University of Genoa and IRCCS San Martino Polyclinic Hospital, Genoa, Italy

Department of Physics of Condensed Matter. University of Seville, Seville, Spain.

Hospital Clinic of Barcelona, Barcelona, Spain

Hospital Regional Universitario Málaga, Málaga, Spain

Methods

DNA of Microbial Keratitis samples was collected with CIM and extracted using the MasterPure Complete DNA and RNA Purification Kit (Epicentre). DNA was fragmented by sonication into fragments of 300 to 400 base pairs (bp) using Bioruptor (Diagenode, Belgium) and then used as a template for library preparation. DNA libraries were sequenced on Illumina HiSeq2500. The resulting reads were quality controlled, trimmed and mapped against the human reference genome. The unmapped reads were taxonomically classified using the Kraken software.

Results

18 microbial keratitis samples were included in the study. Brevundimonas diminuita was found in 5 samples while 6 samples showed the presence of viral infections. Cutibacterium acnes, Staphylococcus aureus, Moraxella lacunata and Pseudomonas alcaligenes were also identified as the presumed putative cause of the infection.

Conclusions

Shotgun sequencing can be used as a diagnostic tool in microbial keratitis samples. This diagnostic method expands the available tests to diagnose eye infections and could be clinically significant in culture negative samples.