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A novel fluorescent assay based on DNAzyme-assisted detection of prostate specific antigen for signal amplification
Analytica Chimica Acta ( IF 6.2 ) Pub Date : 2020-04-01 , DOI: 10.1016/j.aca.2020.01.014
Ying Yan 1 , Changbei Ma 1 , Zhenwei Tang 1 , Mingjian Chen 1 , Han Zhao 1
Affiliation  

Prostate specific antigen (PSA) is one of the most common biomarkers for the management of prostate cancer. However, it still remains urgent to develop highly sensitive, cost-effective and selective strategies for PSA assay. In this paper, we developed a low-cost, highly sensitive and specific analytical strategy for the detection of PSA by using a fluorescence sensor based on Pb2+-dependent DNAzyme. We designed a DNA sequence called cmMB with a hairpin structure, containing PSA-specific aptamers and Pb2+-dependent DNAzyme chains. Also, a fluorophore-labelled DNA sequence called Sub-FAM, which contains a cleavage site of Pb2+-dependent DNAzyme and serves as substrate, is also designed for the signal generation. In the presence of PSA, interaction between aptamer and PSA blocks the hairpin structure of cmMB, resulting in the formation of Pb2+-dependent DNAzyme with Pb2+. Then, Pb2+-dependent DNAzyme can cleavage Sub-FAM and produce a high fluorescence. In the absence of PSA, since Sub-FAM remains to be ssDNA and can be absorbed by GO, only low fluorescence can be detected. Under optimal experimental conditions, a good linear relationship in the range of 1-100 pg mL-1 was exhibited, with a limit of detection (LOD) of 0.76 pg mL-1. In addition, the proposed method has potential value in the diagnosis and monitoring of prostate cancer because of its good selectivity and practical application in biological samples.

中文翻译:

基于 DNA 酶辅助检测前列腺特异性抗原的新型荧光检测用于信号放大

前列腺特异性抗原 (PSA) 是治疗前列腺癌最常见的生物标志物之一。然而,开发用于 PSA 检测的高度灵敏、具有成本效益和选择性的策略仍然很紧迫。在本文中,我们开发了一种低成本、高灵敏度和特异性的分析策略,通过使用基于 Pb2+ 依赖性 DNAzyme 的荧光传感器检测 PSA。我们设计了一个名为 cmMB 的 DNA 序列,具有发夹结构,包含 PSA 特异性适体和 Pb2+ 依赖性 DNAzyme 链。此外,荧光团标记的 DNA 序列称为 Sub-FAM,它包含 Pb2+ 依赖性 DNAzyme 的切割位点并用作底物,也被设计用于信号生成。在 PSA 存在下,适配体和 PSA 之间的相互作用阻断了 cmMB 的发夹结构,导致与 Pb2+ 形成依赖于 Pb2+ 的 DNAzyme。然后,依赖 Pb2+ 的 DNAzyme 可以切割 Sub-FAM 并产生高荧光。在没有 PSA 的情况下,由于 Sub-FAM 仍然是 ssDNA 并且可以被 GO 吸收,因此只能检测到低荧光。在最佳实验条件下,在 1-100 pg mL-1 范围内表现出良好的线性关系,检测限 (LOD) 为 0.76 pg mL-1。此外,该方法具有良好的选择性和在生物样品中的实际应用,在前列腺癌的诊断和监测中具有潜在价值。在最佳实验条件下,在 1-100 pg mL-1 范围内表现出良好的线性关系,检测限 (LOD) 为 0.76 pg mL-1。此外,该方法具有良好的选择性和在生物样品中的实际应用,在前列腺癌的诊断和监测中具有潜在价值。在最佳实验条件下,在 1-100 pg mL-1 范围内表现出良好的线性关系,检测限 (LOD) 为 0.76 pg mL-1。此外,该方法具有良好的选择性和在生物样品中的实际应用,在前列腺癌的诊断和监测中具有潜在价值。
更新日期:2020-04-01
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