ArticleBMC research notes2017
Development and validation of a real-time PCR assay for the detection of clinical acanthamoebae.
Article in BMC research notes, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 40 citations in OpenAlex.
- Article
- Article
- Nested PCR as a superior diagnostic method for Acanthamoeba keratitis compared to conventional techniques.Scientific reports · 2026Article
- Incidence of Acanthamoeba Keratitis in Switzerland.Microorganisms · 2025Article
- EnhancingMicrobiology spectrum · 2025Article
- Assessing PCR-Positive Acanthamoeba Keratitis-A Retrospective Chart Review.Microorganisms · 2024Article
- Chitosan nanoparticles improve the effectivity of miltefosine against Acanthamoeba.PLoS neglected tropical diseases · 2024Article
- New Frontiers inBiology · 2023Review
- Diagnosis ofDiagnostics (Basel, Switzerland) · 2023Review
- Impact of implementation of polymerase chain reaction on diagnosis, treatment, and clinical course of Acanthamoeba keratitis.Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie · 2023Article
- Real-Time PCR for the Diagnosis ofMicroorganisms · 2022Article
- Surveillance of Amoebic Keratitis-Causing Acanthamoebae for Potential Bacterial Endosymbionts in Ontario, Canada.Pathogens (Basel, Switzerland) · 2022Article
- Diagnostic methods for the etiological assessment of infectious corneal pathology (Review).Experimental and therapeutic medicine · 2022Review
- Diagnostic armamentarium of infectious keratitis: A comprehensive review.The ocular surface · 2022Review
- Efficient nested-PCR-based method development for detection and genotype identification of Acanthamoeba from a small volume of aquatic environmental sample.Scientific reports · 2021Article
Corrections and comments
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Authors and funding
10 authors at 5 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundSuboptimal agreement between molecular assays for the detection of Acanthamoeba spp. in clinical specimens has been demonstrated, and poor assay sensitivity directly imperils the vision of those affected by amoebic keratitis (AK) through delayed diagnosis. We sought to develop and validate a single Taqman real time PCR assay targeting the Acanthamoeba 18S rRNA gene that could be used to enhance sensitivity and specificity when paired with reference assays.
methodsBiobanked DNA from surplus delinked AK clinical specimens and 10 ATCC strains of Acanthamoeba was extracted. Sequence alignment of 66 18S rRNA regions from 12 species of Acanthamoeba known to cause keratitis informed design of a new TaqMan primer set. Performance of the new assay was compared to the 2 assays used currently in our laboratory.
resultsAmong 24 Acanthamoeba-positive and 83 negative specimens by the CDC reference standard, performance characteristics of the newly designed primer set were as follows: sensitivity 100%, specificity 94%, PPV 82.8%, and NPV 100%. Compared to culture, sensitivity of the new primer set was 100%, and specificity 96%. No cross-reactivity of the primer set to non-acanthamoebae, including Balamuthia and Naegleria, was found.
conclusionsWe have validated a real time PCR assay for the diagnosis of AK, and in doing so, have overcome important barriers to rapid and sensitive detection of acanthamoebae, including limited sensitivity and specificity of commonly used assays.
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