Jahanpeyma Fatemeh, Forouzandeh Mehdi, Rasaee Mohammad Javad, Shoaie Nahid
Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran,
Front Biosci (Schol Ed). 2019 Mar 1;11(1):122-135. doi: 10.2741/S530.
Point-of-care Nucleic acid testing (POCNAT) has become an attractive technique for DNA identification in resource-limited settings, offering a rapid system for urgent clinical applications. In this study, a chemiluminescence-based lateral flow biosensor (CL-LFB) was developed for the quantitative analysis of DNA, without labeling and amplification. The developed biosensor employs a two-step hybridization, a primary hybridization of 5'-biotinylated detector probe to the target DNA and a secondary hybridization of the resulting complex with the immobilized capture probe. Quantitative analysis of DNA was provided via HRP-catalyzed reaction with the chemiluminescense substrate followed by imaging with a complementary metal-oxide-semiconductor (CMOS) digital camera. The assay performance was investigated using a synthetic target, gene (775 bp) and the whole genome derived from A detection limit of 1.5 pM for the synthetic target and 0.4 ng/ml for gene was obtained. In spite of LFBs limitations for the detection of large DNA fragments, the proposed assay provided a low-cost, fast, and sensitive tool for PCR-free diagnosis of small and larger fragments of nucleic acids.
即时核酸检测（POCNAT）已成为资源有限环境中用于DNA鉴定的一种有吸引力的技术，为紧急临床应用提供了一种快速系统。在本研究中，开发了一种基于化学发光的侧向流动生物传感器（CL-LFB）用于DNA的定量分析，无需标记和扩增。所开发的生物传感器采用两步杂交，第一步是将5'-生物素化的检测探针与目标DNA进行初次杂交，第二步是将所得复合物与固定化的捕获探针进行二次杂交。通过HRP催化与化学发光底物的反应，然后用互补金属氧化物半导体（CMOS）数码相机成像，对DNA进行定量分析。使用合成靶标、基因（775 bp）和源自的全基因组研究了该检测性能。合成靶标的检测限为1.5 pM，基因的检测限为0.4 ng/ml。尽管侧向流动生物传感器在检测大片段DNA方面存在局限性，但所提出的检测方法为无PCR诊断小片段和大片段核酸提供了一种低成本、快速且灵敏的工具。
