Hanith Raj Deivarajan, Prabhakara SethupathyR, Vignesh Elamurugan, Akshayaa Vs, Reega P, Dharani Chelliah, Hari Vignesh S, Elakkiya Nandhini Gr, Kanmani M, Dharsini N, Karvannan Sevugamurthi, Saravanan Vr, Anuradha K, Parag K Shah, Ram Rammohan, Abhishek Nair, Krutin Shah, Anju Jose, Jaishree Pandian, Lalitha Prajna, Venkatesh N Prajna, Siddharth Narendran
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引用次数: 0
Abstract
Objective: This study evaluates the RID-MyC (Rapid Identification of Mycoses using CRISPR) assay, a CRISPR/Cas12a-based diagnostic tool, for its efficacy in diagnosing fungal endophthalmitis (FE), comparing it with panfungal PCR and culture methods.
Design: A comparative cross-sectional study assessing the performance of the RID-MyC assay against established diagnostic modalities for FE.
Subjects: The study included 133 intraocular samples from 117 patients with suspected microbial endophthalmitis.
Methods: The study compared the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the RID-MyC assay against panfungal PCR and culture. The Limit of Detection (LoD) for Aspergillus flavus and Candida albicans was determined for both RID-MyC and panfungal PCR across three different media: nuclease-free water (NFW), aqueous humor (AH), and vitreous humor (VH). Discrepancy analysis was conducted for discordant results, incorporating clinical outcomes and responses to antifungal treatment.
Main outcome measures: The study primarily assessed the sensitivity, specificity, PPV, and NPV for clinical samples. Time to diagnosis was also evaluated.
Results: The RID-MyC assay demonstrated a sensitivity of 88.24% (CI: 63.56% to 98.54%) and specificity of 93.1% (CI: 86.86% to 96.98%), with PPV and NPV of 65.22% (CI: 48.45% to 78.91%) and 98.18% (CI: 93.62% to 99.50%), respectively. Discrepancy analysis enhanced sensitivity to 90.48% (CI: 69.62% to 98.83%) and specificity to 96.43% (CI: 91.11% to 99.02%). The RID-MyC assay was 10 to 1000-fold more sensitive than panfungal PCR in detecting Aspergillus flavus and Candida albicans in intraocular specimens. The time to diagnosis with the RID-MyC assay was consistently under two hours.
Conclusions: The RID-MyC assay may advance the rapid and precise diagnosis of FE, with possible relevance to other invasive fungal conditions.
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