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电化学芯片基基因磁珠法检测高危型人乳头瘤病毒 DNA。

Electrochemical chip-based genomagnetic assay for detection of high-risk human papillomavirus DNA.

机构信息

Regional Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53 Brno, Czech Republic.

Regional Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53 Brno, Czech Republic.

出版信息

Biosens Bioelectron. 2016 Sep 15;83:300-5. doi: 10.1016/j.bios.2016.04.035. Epub 2016 Apr 14.

Abstract

Cervical cancer, being the fourth leading cause of cancer death in women worldwide, predominantly originates from a persistent infection with a high-risk human papillomavirus (HPV). Detection of DNA sequences from these high-risk strains, mostly HPV-16 and HPV-18, represents promising strategy for early screening, which would help to identify women with higher risk of cervical cancer. In developing countries, inadequate screening options lead to disproportionately high mortality rates, making a fast and inexpensive detection schemes highly important. Electrochemical sensors and assays offer an alternative to current methods of detection. We developed an electrochemical-chip based assay, in which target HPV DNA is captured via magnetic bead-modified DNA probes, followed by an antidigoxigenin-peroxidase detection system at screen-printed carbon electrode chips, enabling parallel measurements of eight samples simultaneously. We show sensitive detection in attomoles of HPV DNA, selective discrimination between HPV-16 and HPV-18 and good reproducibility. Most importantly, we show application of the assay into both cancer cell lines and cervical smears from patients. The electrochemical results correlated well with standard methods, making this assay potentially applicable in clinical practice.

摘要

宫颈癌是全球女性癌症死亡的第四大主要原因,主要源于高危型人乳头瘤病毒(HPV)的持续感染。检测这些高危型 HPV-16 和 HPV-18 等病毒的 DNA 序列,是早期筛查的一种有前景的策略,有助于识别出宫颈癌风险较高的女性。在发展中国家,由于筛查手段不足,导致死亡率不成比例地升高,因此快速且廉价的检测方案显得尤为重要。电化学传感器和检测方法为当前的检测方法提供了替代方案。我们开发了一种基于电化学芯片的检测方法,其中目标 HPV DNA 通过磁珠修饰的 DNA 探针捕获,然后在丝网印刷碳电极芯片上进行抗 digoxigenin-过氧化物酶检测系统,从而能够同时平行测量 8 个样本。我们展示了对 HPV DNA 的纳摩尔级灵敏检测、HPV-16 和 HPV-18 的选择性区分以及良好的重现性。最重要的是,我们展示了该检测方法在癌细胞系和患者宫颈涂片上的应用。电化学检测结果与标准方法高度相关,这使得该检测方法在临床实践中具有潜在的应用价值。

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