ArticleAmerican journal of ophthalmology case reports2020
Shotgun sequencing to determine corneal infection.
Article in American journal of ophthalmology case reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
12 citing papers in PubMed, 22 citations in OpenAlex.
- Contact lens-associated infectious keratitis in physicians: a retrospective observational study.International ophthalmology · 2026Observational
- The gut-eye axis in blinding eye diseases: microbiota-driven immune dysregulation and immunomodulatory therapies.International ophthalmology · 2025Review
- Pseudomonas aeruginosa keratitis in a tertiary referral center: a decade of clinical insights and therapeutic challenges.Journal of ophthalmic inflammation and infection · 2025Article
- Enhanced Detection of Bacterial Ocular Pathogens: A Comparative Study of Broad-Range Real-Time PCR and Conventional Culture Methods.Diagnostics (Basel, Switzerland) · 2025Article
- A Comparative Analysis of the Ocular Microbiome: Insights into Healthy Eyes and Anophthalmic Sockets.Microorganisms · 2024Article
- The immunobiology of corneal HSV-1 infection and herpetic stromal keratitis.Clinical microbiology reviews · 2024Review
- Two-stage deep neural network for diagnosing fungal keratitis via in vivo confocal microscopy images.Scientific reports · 2024Article
- Clinical applications of metagenomics next-generation sequencing in infectious diseases.Journal of Zhejiang University. Science. B · 2024Review
- Herpes simplex keratitis: A brief clinical overview.World journal of virology · 2024Review
- Herpetic Keratitis Following Corneal Crosslinking for Keratoconus: A Case Series.Infection and drug resistance · 2022Article
- Metagenomic nanopore sequencing of ocular microbiome in patients with meibomian gland dysfunction.Frontiers in medicine · 2022Article
- Metagenome techniques to reduce diagnostic delay in Acanthamoeba keratitis.Romanian journal of ophthalmologyArticle
Corrections and comments
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Authors and funding
10 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
purposeTo investigate if shotgun-sequencing method could be useful in detailed diagnosis of herpes simplex virus (HSV) infection and compare it with the conventional diagnostic method. OBSERVATIONS: Using a sterile scraper, the infectious part of the ocular surface was scraped gently and placed on a glass slide for conventional diagnosis using PCR and histology and in RNA stabilizing reagent for shotgun sequencing respectively. Concentration of the DNA was determined using a sensitive fluorescence dye-based Qubit dsDNA HS Assay Kit. Shotgun-sequencing libraries were generated using the NEBNext DNA ultra II protocol. The samples were sequenced on the Illumina NextSeq 500 in high output mode with 2X150 bp paired-end sequencing. Taxonomic and functional profiles were generated.Conventional diagnostic method suspected herpetic keratitis. The results indicated presence of an amplified product of 92 bp positive HSV-DNA. Conventional diagnostic method detected the presence of Herpes Simplex Virus DNA (type 1). Shotgun sequencing confirmed the diagnosis of HSV along with the taxonomical profiling of the virus. These results were achieved using 1.9 ng/μL of DNA concentration (114 ng in 60 μL) of the total sample volume. CONCLUSIONS AND IMPORTANCE: Shotgun sequencing is a hypothesis-free approach that identifies full taxonomic and functional profile of an organism. This technology is advantageous as it requires smaller sample size compared to conventional diagnostic methods.
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Registered trials
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