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
摘要
研究目的本研究评估了基于CRISPR/Cas12a的诊断工具RID-MyC（Rapid Identification of Mycoses using CRISPR）测定在诊断真菌性眼内炎（FE）方面的有效性，并将其与泛真菌PCR和培养方法进行了比较：设计：一项横断面比较研究，评估 RID-MyC 检测法与现有 FE 诊断方法的性能：方法：比较 RID-MyC 检测法和现有 FE 诊断方法的灵敏度、特异性和准确性：该研究比较了 RID-MyC 检测法与泛真菌 PCR 和培养法的灵敏度、特异性、阳性预测值 (PPV) 和阴性预测值 (NPV)。研究测定了 RID-MyC 和泛真菌 PCR 在三种不同培养基（无核酸酶水 (NFW)、房水 (AH) 和玻璃体液 (VH)）上对黄曲霉和白色念珠菌的检测限 (LoD)。对不一致的结果进行差异分析，并结合临床结果和对抗真菌治疗的反应：研究主要评估了临床样本的敏感性、特异性、PPV 和 NPV。还评估了诊断时间：RID-MyC测定的灵敏度为88.24%（CI：63.56%至98.54%），特异性为93.1%（CI：86.86%至96.98%），PPV和NPV分别为65.22%（CI：48.45%至78.91%）和98.18%（CI：93.62%至99.50%）。差异分析将灵敏度提高到 90.48%（CI：69.62% 至 98.83%），特异性提高到 96.43%（CI：91.11% 至 99.02%）。在检测眼内标本中的黄曲霉菌和白色念珠菌时，RID-MyC 检测法的灵敏度是泛真菌 PCR 的 10 到 1000 倍。RID-MyC测定的诊断时间始终在两小时以内：结论：RID-MyC 检测法可促进对 FE 的快速、精确诊断，并可能与其他侵袭性真菌疾病相关。
Clinical Evaluation of a Novel CRISPR-Cas12a-Based RID-MyC Assay for the Diagnosis of Fungal Endophthalmitis.
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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