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Clinical Evaluation of a Novel CRISPR-Cas12a-Based RID-MyC Assay for the Diagnosis of Fungal Endophthalmitis
Hanith Raj Deivarajan1, Prabhakara Sethupathy R1, Vignesh Elamurugan2
1Department of Microbiology, Aravind Medical Research Foundation, Madurai, Tamil Nadu, India; Aravind Medical Research Foundation Regional Centre, Coimbatore, Tamil Nadu, India.
Objective:
This study evaluated the RID-MyC (Rapid Identification of Mycoses using clustered regularly interspaced short palindromic repeats [CRISPR]) assay, a CRISPR/Cas12a-based diagnostic tool, for its efficacy in diagnosing fungal endophthalmitis (FE) compared with panfungal polymerase chain reaction (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, and negative predictive value of the RID-MyC assay against panfungal PCR and culture. The limit of detection for Aspergillus flavus and Candida albicans was determined for both RID-MyC and panfungal PCR across 3 different media: nuclease-free water, aqueous humor, and vitreous humor. 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, positive predictive value, and negative predictive value for clinical samples. Time to diagnosis was also evaluated.
Results:
The RID-MyC assay demonstrated a sensitivity of 88.24% (95% confidence interval [CI], 63.56%-98.54%) and specificity of 93.1% (95% CI, 86.86%-96.98%), with positive predictive value and negative predictive value of 65.22% (95% CI, 48.45%-78.91%) and 98.18% (95% CI, 93.62%-99.50%), respectively. Discrepancy analysis enhanced sensitivity to 90.48% (95% CI, 69.62%-98.83%) and specificity to 96.43% (95% CI, 91.11%-99.02%). The RID-MyC assay was 10- to 1000-fold more sensitive than panfungal PCR in detecting A. flavus and C. albicans in intraocular specimens. The time to diagnosis with the RID-MyC assay was consistently <2 hours.
Conclusions:
The RID-MyC assay may advance the rapid and precise diagnosis of FE, with possible relevance to other invasive fungal conditions.
Financial Disclosure(S):
Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.
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