Department of Biochemistry and Biomedical Sciences, McMaster University, 1280 Main Street West, Hamilton, Ontario, L8S 4K1, Canada.
Department of Medicine, McMaster University, 1280 Main Street West, Hamilton, Ontario, L8S 4K1, Canada.
Chemistry. 2023 Jun 27;29(36):e202300240. doi: 10.1002/chem.202300240. Epub 2023 May 16.
Clostridium difficile frequently causes an infectious disease known as Clostridium difficile infection (CDI), and there is an urgent need for the development of more effective rapid diagnostic tests for CDI. Previously we have developed an RNA-cleaving fluorogenic DNAzyme (RFD) probe, named RFD-CD1, that is capable of detecting a specific strain of C. difficile but is too specific to recognize other pathogenic C. difficile strains. To overcome this issue, herein we report RFD-CD2, another RFD that is not only highly specific to C. difficile but also capable of recognizing diverse pathogenic C. difficile strains. Extensive sequence and structure characterization establishes a pseudoknot structure and a significantly minimized sequence for RFD-CD2. As a fluorescent sensor, RFD-CD2 can detect C. difficile at a concentration as low as 100 CFU/mL, thus making this DNAzyme an attractive molecular probe for rapid diagnosis of CDI caused by diverse strains of C. difficile.
艰难梭菌常引起一种名为艰难梭菌感染（CDI）的传染病，因此迫切需要开发更有效的 CDI 快速诊断检测方法。此前，我们开发了一种 RNA 切割荧光 DNA 酶（RFD）探针，命名为 RFD-CD1，它能够检测到一种特定的艰难梭菌菌株，但特异性太高，无法识别其他致病性艰难梭菌菌株。为了解决这个问题，我们在此报告了另一种 RFD，即 RFD-CD2，它不仅对艰难梭菌具有高度特异性，还能够识别多种致病性艰难梭菌菌株。广泛的序列和结构特征分析确定了 RFD-CD2 的假结结构和显著简化的序列。作为荧光传感器，RFD-CD2 可以检测到低至 100 CFU/mL 的艰难梭菌，因此这种 DNA 酶是一种有吸引力的分子探针，可用于快速诊断由多种艰难梭菌菌株引起的 CDI。
