Rapid one-tube RPA-coupled CRISPR/Cas12a-based RID-MyC assay for the diagnosis of fungal keratitis.

IF 1.8 4区医学 Q2 OPHTHALMOLOGY

Indian Journal of Ophthalmology Pub Date : 2025-08-01 Epub Date: 2025-07-28 DOI:10.4103/IJO.IJO_1613_24

Hanith R Deivarajan, Kanmani Senthilkumar, Hari V Sekar, Vignesh Elamurugan, Jaishree Pandian, Anitha Venugopal, Dharmalingam Kuppamuthu, Lalitha Prajna, Venkatesh N Prajna, Siddharth Narendran

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引用次数: 0

Abstract

Purpose: This study introduces and evaluates the single-tube rapid identification of mycoses using CRISPR (ST-RID-MyC) assay. This novel diagnostic tool combines recombinase polymerase amplification (RPA) with CRISPR/Cas12a for the rapid and precise diagnosis of fungal keratitis (FK).

Design: Prospective cross-sectional study.

Methods: Corneal scrapings from 61 patients with suspected microbial keratitis were collected at the Cornea Department of a Tertiary Eye Care Center. The study assessed the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the ST-RID-MyC assay. Additional measures included concordance rates with traditional diagnostic methods and the time to diagnosis.

Results: The ST-RID-MyC assay exhibited a sensitivity of 90% and a specificity of 90.48%, with a PPV of 94.74% and an NPV of 82.61%. The ST-RID-MyC showed substantial agreement with culture and microscopy and perfect concordance with conventional RID-MyC. The mean time to diagnosis was significantly reduced (P < 0.001) using the ST-RID-MyC assay, compared to the traditional RID-MyC assay (6 vs. 32 minutes). Visual assessments demonstrated a high level of inter-observer agreement (kappa = 0.832).

Conclusions: The ST-RID-MyC assay, combining RPA and CRISPR/Cas12a in a single-tube system, offers a rapid, accurate, and resource-efficient diagnostic method for FK, potentially transforming clinical management of this condition by enabling faster therapeutic decisions.

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基于单管rpa偶联CRISPR/ cas12的RID-MyC快速检测诊断真菌性角膜炎

目的：介绍并评价利用CRISPR （ST-RID-MyC）单管快速鉴定真菌病的方法。这种新的诊断工具结合了重组酶聚合酶扩增（RPA）和CRISPR/Cas12a，用于真菌角膜炎（FK）的快速准确诊断。设计:前瞻性横断面研究。方法：收集某三级眼科保健中心角膜科61例疑似细菌性角膜炎患者的角膜刮痕。该研究评估了ST-RID-MyC检测的敏感性、特异性、阳性预测值（PPV）和阴性预测值（NPV）。其他措施包括与传统诊断方法的符合率和诊断时间。结果：ST-RID-MyC检测灵敏度为90%，特异性为90.48%，PPV为94.74%，NPV为82.61%。ST-RID-MyC与培养和镜检结果基本一致，与传统的RID-MyC完全一致。与传统的RID-MyC检测相比，使用ST-RID-MyC检测到诊断的平均时间显著缩短（P < 0.001）（6分钟对32分钟）。视觉评估显示观察者之间的一致性很高（kappa = 0.832）。结论：ST-RID-MyC检测在单管系统中结合RPA和CRISPR/Cas12a，为FK提供了一种快速、准确、资源高效的诊断方法，通过更快的治疗决策，有可能改变这种疾病的临床管理。

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来源期刊

Indian Journal of Ophthalmology OPHTHALMOLOGY-

CiteScore

3.80

自引率

19.40%

发文量

1963

审稿时长

38 weeks

期刊介绍： Indian Journal of Ophthalmology covers clinical, experimental, basic science research and translational research studies related to medical, ethical and social issues in field of ophthalmology and vision science. Articles with clinical interest and implications will be given preference.