Bordeaux Institute in Oncology-BRIC-MoTRIL Team, INSERM U1312, University of Bordeaux, Bordeaux, France.
Department of Biology, ETH Zurich, Zurich, Switzerland.
CRISPR J. 2023 Apr;6(2):140-151. doi: 10.1089/crispr.2022.0070. Epub 2023 Mar 13.
Advances in molecular medicine have placed nucleic acid detection methods at the center of an increasing number of clinical applications. Polymerase chain reaction (PCR)-based diagnostics have been widely adopted for their versatility, specificity, and sensitivity. However, recently reported clustered regularly interspaced short palindromic repeats-based methods have demonstrated equivalent to superior performance, with increased portability and reduced processing time and cost. In this study, we applied Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) technology to the detection of oncogenic rearrangements. We implemented SHERLOCK for the detection of mRNA, a hallmark of chronic myeloid leukemia (CML), and DNA oncogenic alleles, frequently detected in glioblastoma and non-small cell lung cancer (NSCLC). SHERLOCK enabled rapid, sensitive, and variant-specific detection of and alterations. Compared with the gold-standard PCR-based methods currently used in clinic, SHERLOCK achieved equivalent to greater sensitivity, suggesting it could be a new tool in CML and NSCLC, to detect low level of molecular residual disease.
分子医学的进展使核酸检测方法在越来越多的临床应用中处于中心地位。聚合酶链反应（PCR）为基础的诊断因其通用性、特异性和敏感性而被广泛采用。然而，最近报道的基于聚类规则间隔短回文重复（CRISPR）的方法已经证明具有同等甚至更高的性能，具有更高的便携性，并且减少了处理时间和成本。在这项研究中，我们将特异性高灵敏度酶报告解锁（SHERLOCK）技术应用于致癌重排的检测。我们实现了 SHERLOCK 对 mRNA 的检测，这是慢性髓性白血病（CML）的标志，以及经常在胶质母细胞瘤和非小细胞肺癌（NSCLC）中检测到的 DNA 致癌等位基因。SHERLOCK 能够快速、敏感和特异性地检测 和 改变。与目前临床上使用的金标准基于 PCR 的方法相比，SHERLOCK 达到了同等或更高的灵敏度，这表明它可能成为 CML 和 NSCLC 中检测低水平分子残留疾病的新工具。
