School of Public Health, Jilin University, Changchun, Jilin, 130021, PR China.
School of Public Health, Jilin University, Changchun, Jilin, 130021, PR China.
Anal Chim Acta. 2024 Aug 15;1317:342888. doi: 10.1016/j.aca.2024.342888. Epub 2024 Jun 18.
Rapid and sensitive detection of pathogenic bacteria is crucial for disease prevention and control. The CRISPR/Cas12a system with the DNA cleavage capability holds promise in pathogenic bacteria diagnosis. However, the sensitivity of CRISPR-based assays remains a challenge. Herein, we report a versatile and sensitive pathogen sensing platform (HTCas12a) based on the CRISPR/Cas12a system, hybridization chain reaction (HCR) and Poly T-copper fluorescence nanoprobe. The sensitivity is improved by HCR and the Poly-T-Cu reporter probe reduces the overall experiment cost to less than one dollar per sample. Our results demonstrate the specific recognition of target nucleic acid fragments from other pathogens. Furthermore, a good linear correlation between fluorescence intensity and target quantities were achieved with detection limits of 23.36 fM for Target DNA and 4.17 CFU/mL for S.aureus, respectively. The HTCas12a system offers a universal platform for pathogen detection in various fields, including environmental monitoring, clinical diagnosis, and food safety.
快速、灵敏地检测致病菌对于疾病预防和控制至关重要。具有 DNA 切割能力的 CRISPR/Cas12a 系统有望应用于致病菌诊断。然而，基于 CRISPR 的检测方法的灵敏度仍然是一个挑战。本研究报告了一种基于 CRISPR/Cas12a 系统、杂交链式反应（HCR）和 Poly T-铜荧光纳米探针的多功能、灵敏的病原体感应平台（HTCas12a）。HCR 提高了灵敏度，而 Poly-T-Cu 报告探针使每个样本的总实验成本降低到 1 美元以下。研究结果表明，该平台能够特异性识别其他病原体的靶核酸片段。此外，荧光强度与目标数量之间呈现出良好的线性相关性，对靶 DNA 的检测限为 23.36 fM，对金黄色葡萄球菌的检测限为 4.17 CFU/mL。该 HTCas12a 系统为包括环境监测、临床诊断和食品安全在内的各个领域的病原体检测提供了通用平台。
