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用于超灵敏检测DNA的基于荧光碳纳米颗粒的侧向流动生物传感器。

Fluorescent carbon nanoparticle-based lateral flow biosensor for ultrasensitive detection of DNA.

作者信息

Takalkar Sunitha, Baryeh Kwaku, Liu Guodong

机构信息

Department of Chemistry and Biochemistry, North Dakota State University, Fargo, ND 58102, USA.

Department of Chemistry and Biochemistry, North Dakota State University, Fargo, ND 58102, USA.

出版信息

Biosens Bioelectron. 2017 Dec 15;98:147-154. doi: 10.1016/j.bios.2017.06.045. Epub 2017 Jun 23.

Abstract

We report a fluorescent carbon nanoparticle (FCN)-based lateral flow biosensor for ultrasensitive detection of DNA. Fluorescent carbon nanoparticle with a diameter of around 15nm was used as a tag to label a detection DNA probe, which was complementary with the part of target DNA. A capture DNA probe was immobilized on the test zone of the lateral flow biosensor. Sandwich-type hybridization reactions among the FCN-labeled DNA probe, target DNA and capture DNA probe were performed on the lateral flow biosensor. In the presence of target DNA, FCNs were captured on the test zone of the biosensor and the fluorescent intensity of the captured FCNs was measured with a portable fluorescent reader. After systematic optimizations of experimental parameters (the components of running buffers, the concentration of detection DNA probe used in the preparation of FCN-DNA conjugates, the amount of FCN-DNA dispensed on the conjugate pad and the dispensing cycles of the capture DNA probes on the test-zone), the biosensor could detect a minimum concentration of 0.4 fM DNA. This study provides a rapid and low-cost approach for DNA detection with high sensitivity, showing great promise for clinical application and biomedical diagnosis.

摘要

我们报道了一种基于荧光碳纳米颗粒(FCN)的侧向流动生物传感器,用于超灵敏检测DNA。使用直径约为15nm的荧光碳纳米颗粒作为标签来标记检测DNA探针,该探针与目标DNA的一部分互补。捕获DNA探针固定在侧向流动生物传感器的测试区。在侧向流动生物传感器上进行FCN标记的DNA探针、目标DNA和捕获DNA探针之间的夹心型杂交反应。在存在目标DNA的情况下,FCN被捕获在生物传感器的测试区,并使用便携式荧光阅读器测量捕获的FCN的荧光强度。经过对实验参数(运行缓冲液的成分、制备FCN-DNA缀合物时使用的检测DNA探针的浓度、分配到结合垫上的FCN-DNA的量以及测试区上捕获DNA探针的分配循环)的系统优化后,该生物传感器能够检测到低至0.4 fM的DNA浓度。这项研究提供了一种快速且低成本的高灵敏度DNA检测方法,在临床应用和生物医学诊断方面显示出巨大的潜力。

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