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基于超灵敏电化学 DNA 纳米生物传感器的早期宫颈癌诊断用于实际样本中 HPV-18 的检测。

Early-stage cervical cancer diagnosis based on an ultra-sensitive electrochemical DNA nanobiosensor for HPV-18 detection in real samples.

机构信息

Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran.

Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

出版信息

J Nanobiotechnology. 2020 Jan 13;18(1):11. doi: 10.1186/s12951-020-0577-9.

Abstract

BACKGROUND

In several years ago, infection with human papillomaviruses (HPVs), have been prevalent in the worlds especially HPV type 18, can lead to cervical cancer. Therefore, rapid, accurate, and early diagnosis of HPV for successful treatment is essential. The present study describes the development of a selective and sensitive electrochemical biosensor base on DNA, for early detection of HPV-18. For this purpose, a nanocomposite of reduced graphene oxide (rGO) and multiwalled carbon nanotubes (MWCNTs) were electrodeposited on a screen-printed carbon electrode (SPCE). Then, Au nanoparticles (AuNPs) were dropped on a modified SPCE. Subsequently, single strand DNA (ssDNA) probe was immobilized on the modified electrode. The link attached between AuNPs and probe ssDNA provided by L-cysteine via functionalizing AuNPs (Cys-AuNPs). The differential pulse voltammetry (DPV) assay was also used to electrochemical measurement. The measurement was based on the oxidation signals of anthraquninone-2-sulfonic acid monohydrate sodium salt (AQMS) before and after hybridization between the probe and target DNA.

RESULTS

The calibration curve showed a linear range between 0.01 fM to 0.01 nM with a limit of detection 0.05 fM. The results showed that the optimum concentration for DNA probe was 5 µM. The good performance of the proposed biosensor was achieved through hybridization of DNA probe-modified SPCE with extracted DNA from clinical samples.

CONCLUSIONS

According to the investigated results, this biosensor can be introduced as a proprietary, accurate, sensitive, and rapid diagnostic method of HPV 18 in the polymerase chain reaction (PCR) of real samples.

摘要

背景

在几年前,人乳头瘤病毒(HPV)的感染在世界范围内非常普遍,尤其是 HPV 18 型,可导致宫颈癌。因此,快速、准确、早期诊断 HPV 以进行成功治疗至关重要。本研究描述了基于 DNA 的选择性和灵敏电化学生物传感器的开发,用于早期检测 HPV-18。为此,将还原氧化石墨烯(rGO)和多壁碳纳米管(MWCNT)的纳米复合材料电沉积在丝网印刷碳电极(SPCE)上。然后,将金纳米粒子(AuNPs)滴在修饰的 SPCE 上。随后,将单链 DNA(ssDNA)探针固定在修饰的电极上。通过功能化 AuNPs(Cys-AuNPs),AuNPs 与探针 ssDNA 之间的连接由 L-半胱氨酸提供。差分脉冲伏安法(DPV)也用于电化学测量。该测量基于探针和目标 DNA 杂交前后蒽醌-2-磺酸单水钠盐(AQMS)的氧化信号。

结果

校准曲线显示在 0.01 fM 至 0.01 nM 之间呈线性范围,检测限为 0.05 fM。结果表明,DNA 探针的最佳浓度为 5 µM。通过将 DNA 探针修饰的 SPCE 与从临床样本中提取的 DNA 进行杂交,实现了所提出的生物传感器的良好性能。

结论

根据研究结果,该生物传感器可作为一种专有的、准确的、灵敏的和快速的诊断方法,用于聚合酶链反应(PCR)中对实际样本中的 HPV 18 进行诊断。

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