1. A Multiplex Dot Hybridization Assay For Detection and Differentiation of Acanthamoeba and Herpes Keratitis.
- Author
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Kuo MT, Fang PC, Yu HJ, Chao TL, Chien CC, Chen SH, Wang JR, Tseng SL, Lai YH, Hsiao CC, and Chang TC
- Subjects
- Cornea parasitology, Cornea virology, Diagnosis, Differential, Humans, Limit of Detection, Multiplex Polymerase Chain Reaction, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Acanthamoeba, Acanthamoeba Keratitis diagnosis, Keratitis, Herpetic diagnosis, Nucleic Acid Hybridization methods, Simplexvirus
- Abstract
Purpose: We verified a multiplex dot hybridization (MDH) assay for the rapid detection and differentiation of Acanthamoeba keratitis (AK) and herpes simplex keratitis (HSK)., Methods: Molecular detection of Acanthamoeba and herpes simplex virus in corneal scrapes was performed with the MDH assay and standard diagnostic methods. The experimental group included corneal scrapes (n = 33) from patients with culture- or pathology-confirmed AK (n = 15) and real-time PCR-confirmed HSK (n = 16). The control group included 50 samples from cases of bacterial keratitis (n = 15), fungal keratitis (n = 15), and initially presumed AK (n = 5) or HSK (n = 17) which finally were excluded by culture for Acanthamoeba or by real-time PCR for herpes simplex virus, respectively. Discrepant results between methods were resolved by DNA sequencing of the PCR amplicons., Results: After discrepant analysis, the sensitivity for the diagnosis of AK and HSK was both 93.3% by the MDH assay, while the specificity was 100% for the two types of keratitis. The turnaround time of MDH assay was within a working day using an already prepared array. Two false-negatives (one AK case and one HSK case) were obtained by the MDH assay., Conclusions: The MDH assay could effectively prevent missed or delayed diagnosis of AK and HSK and has a potential to be adopted in routine clinical practice if the test is commercialized.
- Published
- 2016
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