Purpose: Current diagnostic workup for infectious uveitis relies on the use of various immunoassays and PCR tests on ocular fluids that are time consuming and not comprehensive, requiring large sample volumes to probe for all causes. To streamline the diagnosis process, improve turnaround time and target inclusivity, we developed a highly multiplexed pan-ocular-pathogen panel (OcuPan) that can be used for broad-range pathogen detection in only one assay, with PCR-level sensitivity and in a timely fashion (12h). Here, we assessed the clinical usefulness and performance of this innovative assay as a novel tool for rapid and accurate laboratory diagnosis of infectious uveitis.
Design: Laboratory evaluation of a novel diagnostic test and technology PARTICIPANTS AND CONTROLS: A total of 109 patients from two centers were included, including 78 infectious uveitis cases and 31 controls.
Methods: Intraocular samples (n=108 from patients and 43 from controls) were processed by PCR and the OcuPan diagnostic assay that detects 46 ocular pathogens plus two resistance/virulence markers.
Main outcome measures: Clinical sensitivity and specificity, quantification capabilities, and agreement between PCR and the OcuPan assay.
Results: The clinical sensitivity and specificity of the OcuPan assay were 50.9% (95% confidence interval [CI]), 41.3 to 60.5) and 100% (95% CI, 89.7% to 100%), respectively. Sensitivity varied according to the sample matrix tested, with undiluted vitreous having the highest positivity rates (72.4%; 95% CI, 55.1 to 89.7), followed by dilute vitreous (53.3%; 95% CI, 34.4 to 72.3) and aqueous (36.7%; 95% CI, 22.7 to 50.7). There was excellent overall agreement (93.5%) between the OcuPan assay and PCR (kappa of 0.870, ±0.047; P <0.001) with positive (PPA) and negative (NPA) percent agreements >92% for all targets. PPA was 100% for herpesviruses and T. pallidum, while the NPA values ranged from 96.6% to 100% for these pathogens. For T. gondii the PPA was 75% and NPA 100%. We also found significant correlation (Spearman rho= -0.7506, P <0.0001) between the quantitative metric for the OcuPan assay and the real time PCR Ct values.
Conclusions: The OcuPan assay offers an all-in-one highly multiplexed detection system for rapid, comprehensive, quantitative and accurate diagnosis of infectious uveitis.
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