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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
PCR and culture for diagnosis of Acanthamoeba keratitis
British Journal of Ophthalmology, Volume 105, No. 9, Year 2021
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Description
Background/Aims Acanthamoeba keratitis (AK) is a rare but sight-threatening infection. Molecular diagnosis of corneal scraping has improved the diagnosis of AK. Different molecular targets and conditions have been used in diagnosis thus far. In this study, we prospectively compared the performance of five PCR assays on corneal samples for the diagnosis of AK. Methods 1217 corneal scraping samples were obtained from patients, for whom an AK was suspected. Sample processing involved both molecular diagnostics and culture. Acanthamoeba PCR assays detected different regions of the Acanthamoeba nuclear small-subunit rRNA gene: three final point PCR assays using Nelson, ACARNA and JDP1-JDP2 pairs of primers, and two real-time PCR assays using Acant primer-probe. Human DNA and internal control were co-amplified in the real-time PCR assay to ensure scraping quality and the absence of inhibitors. In the absence of a gold standard, the performance of each test was evaluated using latent class analysis. Genotypes of Acanthamoeba isolates were also characterised. Results Estimated prevalence of AK was 1.32%. The sensitivity of Acanthamoeba diagnostic PCRs (73.3% to 86.7%) did not differ significantly from that of culture (66.7%), or according to the target sequence or the technology. Sensitivity could be increased to 93.8% or 100% by combining two or three assays, respectively. PCR specificity (99.3% to 100%) differed between the assays. T4 was the predominant Acanthamoeba genotype (84.6%). Conclusions Culture and a single PCR assay could lead to misdiagnosing AK. A combination of different PCR assays and improved sample quality could increase diagnosis sensitivity. © 2021 BMJ Publishing Group. All rights reserved.
Authors & Co-Authors
Yéra, Hélène Ne
France, Paris
Université Paris Cité
France, Paris
Hopital Cochin Ap-hp
Ariey, Frédéric
France, Paris
Université Paris Cité
France, Paris
Hopital Cochin Ap-hp
Hasseine, Lilia
France, Nice
Centre Hospitalier Universitaire de Nice
Delaunay, Pascal
France, Nice
Centre Hospitalier Universitaire de Nice
Marty, Pierre M.
France, Nice
Centre Hospitalier Universitaire de Nice
France, Nice
Université Côte D'azur
Statistics
Citations: 24
Authors: 5
Affiliations: 4
Identifiers
Doi:
10.1136/bjophthalmol-2020-316730
ISSN:
00071161
Research Areas
Cancer
Genetics And Genomics
Study Design
Cross Sectional Study