{"title":"Clinical Evaluation of a Novel CRISPR-Cas12a-Based RID-MyC Assay for the Diagnosis of Fungal Endophthalmitis.","authors":"Hanith Raj Deivarajan, Prabhakara Sethupathy R, Vignesh Elamurugan, Akshayaa Vs, Reega P, Dharani Chelliah, Hari Vignesh S, Elakkiya Nandhini Gr, Kanmani M, Dharsini Nandhakumar, 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","doi":"10.1016/j.oret.2024.11.003","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>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.</p><p><strong>Design: </strong>A comparative cross-sectional study assessing the performance of the RID-MyC assay against established diagnostic modalities for FE.</p><p><strong>Subjects: </strong>The study included 133 intraocular samples from 117 patients with suspected microbial endophthalmitis.</p><p><strong>Methods: </strong>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.</p><p><strong>Main outcome measures: </strong>The study primarily assessed the sensitivity, specificity, positive predictive value, and negative predictive value for clinical samples. Time to diagnosis was also evaluated.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>The RID-MyC assay may advance the rapid and precise diagnosis of FE, with possible relevance to other invasive fungal conditions.</p><p><strong>Financial disclosure(s): </strong>Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.</p>","PeriodicalId":19501,"journal":{"name":"Ophthalmology. Retina","volume":" ","pages":""},"PeriodicalIF":4.4000,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Ophthalmology. Retina","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.oret.2024.11.003","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"OPHTHALMOLOGY","Score":null,"Total":0}
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Abstract
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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