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Next-generation sequencing for the detection of microorganisms present in human donor corneal preservation medium

Mohit Parekh1,2, Davide Borroni3, Vito Romano3,4,5

  • 1Institute of Ophthalmology, University College London, London, UK.

BMJ Open Ophthalmology
|June 11, 2019
PubMed
Abstract

Insights

Next-generation sequencing detected various bacteria and fungi in cornea storage media, unlike conventional methods. This metagenomic approach offers broader microbiological insights for clinical practice.

Area of Science:

  • Microbiology
  • Genomics
  • Ophthalmology

Background:

  • Human donor corneas require sterile storage media for transplantation.
  • Current diagnostic methods may not detect all microorganisms present in storage media.

Purpose of the Study:

  • To detect microorganisms in human donor cornea storage media using next-generation sequencing (NGS).
  • To compare NGS with conventional diagnostic methods (CDM).

Main Methods:

  • Samples from organ culture (OC) and hypothermic storage media were analyzed.
  • Ribosomal RNA sequencing (16S and 18S) was performed.
  • Results were compared with conventional diagnostic methods (Bactec).

Main Results:

  • NGS identified abundant bacterial genera including *Pseudomonas* and *Comamonas* in both storage types.
  • Fungal genus *Malassezia* was detected in both storage conditions.
  • CDM yielded negative results for all samples, indicating limitations in detecting low microbial loads.

Conclusions:

  • Metagenomics offers comprehensive taxonomic profiling for broader microbiological diagnostics.
  • NGS shows potential for clinical application in cornea storage media analysis.
  • Reducing costs and turnaround time, and detecting viable organisms are key for routine clinical implementation.
Keywords:
16S18SNGSbacteriacorneaeye bankfungusmediamicrobiologypreservationstorage

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