College of Bioresources Chemical and Materials Engineering, Shaanxi University of Science & Technology, Xi'an 710021, P. R. China.
National Demonstration Center for Experimental Light Chemistry Engineering Education, Shaanxi University of Science & Technology, Xi'an 710021, P. R. China.
Anal Methods. 2023 Jul 6;15(26):3240-3250. doi: 10.1039/d3ay00695f.
Nucleic acid detection technologies have been widely utilized for various diseases. Conventional laboratory tests are less suitable for use in resource-limited settings as they are time-consuming, high-cost, complex, and heavily dependent on benchtop equipment. Rapid nucleic acid detection methods that consist of rapid nucleic acid extraction steps could overcome these challenges. A paper-based platform has been utilized to develop various rapid nucleic acid extraction methods owing to its cost-effectiveness, portability, and easy-modification. However, the existing paper-based nucleic acid extraction technologies mainly focus on improving the adsorption capacity of nucleic acids without reducing the non-specific adsorption capacity of proteins. In this study, paper-based nucleic acid extraction technology with wash-free, elution-free, and low protein adsorption was developed. The fabrication of paper involves the mixing of polyethylene glycol (PEG)-modified cotton fiber, chitosan (COS)-modified cotton fiber, and cotton fiber to form PEG-modified cotton fiber/chitosan-modified cotton fiber/cotton fiber (PEG-CF/COS-CF/CF) paper by the wet molding method. The result showed that PEG-CF/COS-CF/CF paper has a desirable pore size (23.9 ± 4.03 μm), good mechanical strength (dry: 9.37 Mpa and wet: 0.28 Mpa), and hydrophilicity (contact angle: 42.6° ± 0.36°). NH groups of COS and OH groups of PEG were observed on its surface and the adsorption efficiency of nucleic acid in TE buffer was 42.48% ± 0.30%. The limit of detection of pure DNA with this PEG-CF/COS-CF/CF paper by qPCR was as low as 25 ng. Additionally, this platform could successfully extract nucleic acid from 30 μL of a saliva sample, highlighting its potential use for clinical sample testing. The proposed paper-based nucleic acid extraction platform shows tremendous potential for disease diagnosis in resource-limited settings.
核酸检测技术已广泛应用于各种疾病。传统的实验室检测在资源有限的环境中不太适用，因为它们耗时、成本高、复杂且严重依赖台式设备。快速核酸检测方法包括快速核酸提取步骤，可以克服这些挑战。由于成本效益高、便携性强且易于修改，基于纸张的平台已被用于开发各种快速核酸提取方法。然而，现有的基于纸张的核酸提取技术主要侧重于提高核酸的吸附能力，而不降低蛋白质的非特异性吸附能力。在这项研究中，开发了一种无洗涤、无洗脱且蛋白质吸附能力低的基于纸张的核酸提取技术。纸张的制造涉及将聚乙二醇（PEG）修饰的棉纤维、壳聚糖（COS）修饰的棉纤维和棉纤维混合在一起，通过湿法成型法形成 PEG 修饰的棉纤维/壳聚糖修饰的棉纤维/棉纤维（PEG-CF/COS-CF/CF）纸。结果表明，PEG-CF/COS-CF/CF 纸具有理想的孔径（23.9±4.03μm）、良好的机械强度（干燥：9.37Mpa 和湿：0.28Mpa）和亲水性（接触角：42.6°±0.36°）。其表面观察到 COS 的 NH 基团和 PEG 的 OH 基团，在 TE 缓冲液中核酸的吸附效率为 42.48%±0.30%。使用这种 PEG-CF/COS-CF/CF 纸通过 qPCR 检测纯 DNA 的检测限低至 25ng。此外，该平台可以成功从 30μL 的唾液样本中提取核酸，突出了其在临床样本测试中的潜在用途。该基于纸张的核酸提取平台在资源有限的环境中用于疾病诊断具有巨大潜力。
