Related Experiment Video
Updated: Dec 21, 2025
08:31
Porcine Corneal Tissue Explant to Study the Efficacy of Herpes Simplex Virus-1 Antivirals
Published on: September 20, 2021
3.7K
Shotgun sequencing to determine corneal infection
Mohit Parekh1,2, Vito Romano3,4,5, Antonella Franch2,6
1Institute of Ophthalmology, University College London, London, United Kingdom.
American Journal of Ophthalmology Case Reports
|May 22, 2020
Summary
Shotgun sequencing offers a detailed diagnosis for herpes simplex virus (HSV) infections, identifying the full taxonomic profile. This advanced method requires a smaller sample size than traditional techniques.
Area of Science:
- Ophthalmology
- Virology
- Genomics
Background:
- Herpes simplex virus (HSV) infections, particularly herpetic keratitis, require accurate and timely diagnosis.
- Conventional diagnostic methods like PCR and histology are established but may have limitations in comprehensive profiling.
Observation:
- Ocular surface samples were collected for both conventional diagnostics (PCR, histology) and shotgun sequencing.
- Shotgun sequencing utilized the Illumina NextSeq 500 platform after library preparation with the NEBNext DNA ultra II protocol.
- DNA concentration was quantified using the Qubit dsDNA HS Assay Kit, with successful sequencing achieved at 1.9 ng/μL.
Findings:
- Conventional methods confirmed HSV-DNA (type 1) consistent with herpetic keratitis.
- Shotgun sequencing not only confirmed the HSV diagnosis but also provided detailed taxonomic profiling of the virus.
- The hypothesis-free nature of shotgun sequencing allowed for a comprehensive understanding of the viral profile.
Implications:
- Shotgun sequencing provides a hypothesis-free, in-depth diagnostic approach for viral infections like HSV.
- This genomic technology is advantageous due to its ability to yield full taxonomic and functional profiles from smaller sample volumes.
- Shotgun sequencing represents a significant advancement in the detailed diagnosis of ocular viral infections.
