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- Title
Shotgun metagenomic sequencing in culture negative microbial keratitis.
- Authors
Borroni, Davide; Bonzano, Chiara; Sánchez-González, José-María; Rachwani-Anil, Rahul; Zamorano-Martín, Francisco; Pereza-Nieves, Jorge; Traverso, Carlo Enrico; García Lorente, María; Rodríguez-Calvo-de-Mora, Marina; Esposito, Alfonso; Godin, Fernando; Rocha-de-Lossada, Carlos
- Abstract
Purpose: To evaluate the microbiota of culture negative Corneal Impression Membrane (CIM) microbial keratitis samples with the use of shotgun metagenomics analysis. Methods: DNA of microbial keratitis samples were 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 diminuta 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 in 7 samples. 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.
- Publication
European Journal of Ophthalmology, 2023, Vol 33, Issue 4, p1589
- ISSN
1120-6721
- Publication type
Article
- DOI
10.1177/11206721221149077