Rapid and Sensitive Diagnosis of Leber Hereditary Optic Neuropathy Variants Using Detection

文献类型： 外文期刊

第一作者： Wan, Xiaoling

作者： Wan, Xiaoling;Chen, Jieqiong;Wu, Yidong;Chen, Zhixuan;Li, Tong;Sun, Junran;Zhang, Ting;Wang, Xinjie;Sun, Xiaodong;Wan, Xiaoling;Chen, Jieqiong;Wu, Yidong;Chen, Zhixuan;Liu, Yin;Li, Tong;Sun, Junran;Wang, Xinjie;Sun, Xiaodong;Wan, Xiaoling;Chen, Jieqiong;Wu, Yidong;Chen, Zhixuan;Liu, Yin;Li, Tong;Sun, Junran;Zhang, Ting;Li, Yang;Sun, Xiaodong;Liu, Yin;Zhou, Fuling;Li, Yang;Li, Tong;Sun, Junran;Zhang, Ting;Li, Yang;Sun, Xiaodong;Wang, Xinjie;Sun, Xiaodong

作者机构：

期刊名称：JOURNAL OF MOLECULAR DIAGNOSTICS （ 影响因子：4.1； 五年影响因子：4.9 ）

ISSN： 1525-1578

年卷期： 2023 年 25 卷 8 期

页码：

收录情况： SCI

摘要： Leber hereditary optic neuropathy (LHON) is the most common maternally inherited mitochondrial disease, with >90% of cases harboring one of three point variants (m.3460G>A, m.11778G>A, and m.14484T>C). Rapid and sensitive diagnosis of LHON variants is urgently needed for early diagnosis and timely treatment after onset, which is currently limited. Herein, we adapted the Cas12a-based DNA detection platform for LHON mitochondrial variant diagnosis. Single-strand guide CRISPR RNAs and enzymatic recombinase amplification primers were first screened, the CRISPR/Cas12a system was then optimized with restriction enzymes, and finally compared with Sanger sequencing and next -generation sequencing (NGS) in multicenter clinical samples. This approach can be completed within 30 minutes using only one drop of blood and could reach a sensitivity of 1% of heteroplasmy. Among the 182 multicenter clinical samples, the CRISPR/Cas12a detection system showed high consistency with Sanger sequencing and NGS in both specificity and sensitivity. Notably, a sample harboring a de novo 3.78% m.11778G>A variant detected by NGS, but not by Sanger sequencing, was successfully confirmed using the CRISPR/Cas12a assay, which proved the effectiveness of our method. Overall, our CRISPR/Cas12a detection system provides an alternative for rapid, convenient, and sensitive detection of LHON variants, exhibiting great potential for clinical practice. (J Mol Diagn 2023

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