Article Text
Abstract
Objective 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 16 S and 18 S. 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 turn-around time need to be reduced, and; the detection of viable organisms would help this technology to be introduced into routine clinical practice.
- cornea
- eye bank
- microbiology
- NGS
- 16S
- 18S
- bacteria
- fungus
- storage
- preservation
- media
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Footnotes
MP and DB contributed equally.
Contributors Concept and design of the study: MP, DB, DC, SF. Drafting manuscript: MP, DB, SBK, SF. Critical revision of manuscript: MP, DB, VR, SBK, DC, DP, SF. Supervision: SBK, DC, DP, SF. Final approval: MP, DB, VR, SBK, DC, DP, SF.
Funding 2017 Camera di Commercio di Venezia Rovigo Delta Lagunare funds. CUP I78C17000040005. 5X1000 funds (year 2016) to Fondazione Banca degli Occhi del Veneto Onlus (Venice, Italy) by the Italian Ministry of Health and Italian Ministry of University and Research.
Competing interests None declared.
Patient consent for publication Not required.
Provenance and peer review Not commissioned; externally peer reviewed.