An All-in-One Highly Multiplexed Diagnostic Assay for Rapid, Sensitive, and Comprehensive Detection of Intraocular Pathogens.

Humans Sensitivity and Specificity Endophthalmitis / diagnosis Bacteria / genetics Polymerase Chain Reaction Eye Infections

Journal

American journal of ophthalmology
ISSN: 1879-1891
Titre abrégé: Am J Ophthalmol
Pays: United States
ID NLM: 0370500

Informations de publication

Date de publication:
06 2023
Historique:
received: 26 08 2022
revised: 08 01 2023
accepted: 17 01 2023
medline: 12 5 2023
pubmed: 29 1 2023
entrez: 28 1 2023
Statut: ppublish

Résumé

Intraocular infections are sight-threatening conditions that can lead to vision loss. Rapid identification of the etiologies plays a key role in early initiation of effective therapy to save vision. However, current diagnostic modalities are time consuming and lack sensitivity and inclusiveness. We present here a newly developed comprehensive ocular panel designed to improve diagnostic yields and provide a tool for rapid and sensitive pathogen detection. Experimental laboratory investigation. A panel containing 46 pathogens and 2 resistance/virulence markers that are commonly detected in intraocular infections was developed. Genomic targets were scrutinized for stretches predicted to be specific for a particular species while being conserved across different strains. A set of primers for sample enrichment, and two 50mer NanoString compatible probes were then designed for each target. Probe-target hybrids were detected and quantified using the NanoString nCounter SPRINT Profiler. Diagnostic feasibility was assessed in a pilot clinical study testing samples from infectious retinitis (n = 15) and endophthalmitis (n = 12) patients, for which the etiologies were confirmed by polymerase chain reaction (PCR) or culture. Analytical studies demonstrated highly sensitive detection of a broad spectrum of pathogens, including bacteria, viruses, and parasites, with limits of detection being as low as 2.5 femtograms per reaction. We also found excellent target specificity, with minimal cross-reactivity detected. The custom-designed NanoString ocular panel correctly identified the causative agent from all clinical specimens positive for a variety of pathogens. This highly multiplexed panel for pathogen detection offers a sensitive, comprehensive, and uniform assay run directly on ocular fluids that could significantly improve diagnostics of sight-threatening intraocular infections.

Identifiants

DOI: 10.1016/j.ajo.2023.01.021 PMID: 36709019
pubmed: 36709019
pii: S0002-9394(23)00030-2
doi: 10.1016/j.ajo.2023.01.021
pii:
doi:

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

82-94

Subventions

Organisme : NEI NIH HHS
ID : R01 EY021558
Pays : United States
Organisme : NEI NIH HHS
ID : R01 EY031600
Pays : United States
Organisme : NEI NIH HHS
ID : R21 EY032231
Pays : United States

Informations de copyright

Copyright © 2023 Elsevier Inc. All rights reserved.

Auteurs

Paulo J M Bispo (PJM)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA; Infectious Disease Institute (P.J.M.B., N.B., J.C., M.S.G., L.S.), Harvard Medical School, Boston, Massachusetts, USA. Electronic address: paulo_bispo@meei.harvard.edu.

Nicole Belanger (N)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA; Infectious Disease Institute (P.J.M.B., N.B., J.C., M.S.G., L.S.), Harvard Medical School, Boston, Massachusetts, USA.

Ashley Li (A)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA.

Renee Liu (R)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA.

Gayatri Susarla (G)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA.

Weilin Chan (W)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA.

James Chodosh (J)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA; Infectious Disease Institute (P.J.M.B., N.B., J.C., M.S.G., L.S.), Harvard Medical School, Boston, Massachusetts, USA.

Michael S Gilmore (MS)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA; Infectious Disease Institute (P.J.M.B., N.B., J.C., M.S.G., L.S.), Harvard Medical School, Boston, Massachusetts, USA; Department of Microbiology and Immunobiology (M.S.G.), Harvard Medical School, Boston, Massachusetts, USA.

Lucia Sobrin (L)

From the Department of Ophthalmology (P.J.M.B., N.B., A.L., R.L., G.S., W.C., J.C., M.S.G., L.S.), Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts, USA; Infectious Disease Institute (P.J.M.B., N.B., J.C., M.S.G., L.S.), Harvard Medical School, Boston, Massachusetts, USA.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › An All-in-One Highly Multiplexed Diagnostic...
questionsmedicales.fr › Environnement et santé publique › Santé publique › Méthodes épidémiologiques › Statistiques comme sujet › Sensibilité et spécificité › An All-in-One Highly Multiplexed Diagnostic...
questionsmedicales.fr › Maladies de l'oeil › Infections de l'oeil › Endophtalmie › An All-in-One Highly Multiplexed Diagnostic...
questionsmedicales.fr › Bactéries › An All-in-One Highly Multiplexed Diagnostic...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Techniques d'amplification d'acides nucléiques › Réaction de polymérisation en chaîne › An All-in-One Highly Multiplexed Diagnostic...
questionsmedicales.fr › Maladies de l'oeil › Infections de l'oeil › An All-in-One Highly Multiplexed Diagnostic...