Qiao Yinuo, Wang Xiaoyan, Kang Xuning, Song Yuzhu, Zhang Jinyang, Han Qinqin
Engineering Research Center for Molecular Diagnosis, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, People's Republic of China.
Anal Methods. 2024 Nov 28;16(46):7927-7939. doi: 10.1039/d4ay01517g.
Tuberculosis is a highly infectious bacterial disease caused by . The spread of this agent has caused serious health problems worldwide, and the rapid and accurate detection of is essential for controlling the spread of infection and for preventing the emergence of multidrug-resistant strains. In this study, the cleavage ability of CRISPR-Cas12a against single-stranded DNA was combined with hybridization chain reaction and chemiluminescent signal to establish an imaging sensor for the hypersensitive detection of DNA. We observed linear relationships between the concentration of DNA and the output signal over the ranges of 10 to 200 pM and 200 to 800 pM DNA. The equations of the standard curves were = 56.08 + 3303, with = 0.9916 for the lower range and = 15.69 + 10 685, with = 0.9929 for the higher range. The limit of detection was as low as 0.83 pM for genomic DNA, and a plasmid containing an -specific sequence was detected at 1 copy per μL. A detection accuracy of 100% was achieved in the analysis of DNA isolated from sputum of hospitalized tuberculosis patients. The sensitivity and specificity of the proposed sensor is combined with a long shelf-life and a low cost of materials. This study introduces a new method for tuberculosis detection and broadens the application of CRISPR-Cas12a-based sensors in clinical diagnosis.
结核病是一种由……引起的高度传染性细菌性疾病。这种病原体的传播在全球范围内引发了严重的健康问题,快速准确地检测……对于控制感染传播和预防多重耐药菌株的出现至关重要。在本研究中,将CRISPR-Cas12a对单链DNA的切割能力与杂交链式反应和化学发光信号相结合,建立了一种用于超灵敏检测……DNA的成像传感器。我们观察到在10至200 pM和200至800 pM DNA浓度范围内,……DNA浓度与输出信号之间存在线性关系。低浓度范围的标准曲线方程为 = 56.08 + 3303,相关系数 = 0.9916,高浓度范围的标准曲线方程为 = 15.69 + 10685,相关系数 = 0.9929。基因组DNA的检测限低至0.83 pM,每微升可检测到1个拷贝的含有……特异性序列的质粒。在对住院结核病患者痰液中分离的DNA进行分析时,检测准确率达到了100%。所提出的传感器的灵敏度和特异性与长保质期和低成本材料相结合。本研究介绍了一种新的结核病检测方法,并拓宽了基于CRISPR-Cas12a的传感器在临床诊断中的应用。 (注:原文中部分关键信息缺失,用“……”表示)