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高产金纳米粒子的人乳头瘤病毒基因分型的 DNA 分离方法从宫颈癌组织样本。

High yield gold nanoparticle-based DNA isolation method for human papillomaviruses genotypes from cervical cancer tissue samples.

机构信息

Department of Medical Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.

Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

IET Nanobiotechnol. 2020 Sep;14(7):555-562. doi: 10.1049/iet-nbt.2020.0093.

DOI:10.1049/iet-nbt.2020.0093
PMID:33010130
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8676591/
Abstract

Gold nanoparticles (AuNPs) are commonly used in biosensors of various kinds. However, its application to extract DNA from cancer tissues has not been extensively studied. The purification of DNA from cancer tissues is an important step in diagnostic and therapeutic development. Almost, all cervical cancer cases are associated with high-risk human papillomavirus (HR-HPV) infection. Accurate viral diagnosis has so far relied on the extraction of adequate amounts of DNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples. Till now, no specific and sensitive DNA purification method has been introduced for the extraction of HR-HPV from FFPE tissue. Since the commercially available purification kits are not sensitive and specific enough for HR-HPV DNA targets, in this study, a DNA purification method was designed based on AuNPs to purify sufficient amounts of HR-HPV DNA from cervical cancer tissue samples. AuNPs were coated with a series of oligonucleotide probes to hybridize to specific DNA sequences of HR-HPV genotypes. Results showed that 733 out of 800 copies of type-specific HPV DNA were recovered with complete specificity, compared to 36 copies with a standard commercial kit (Qiagen FFPE). The high yield of DNA (91.6%) is the main advantage of the AuNPs-probe purification method.

摘要

金纳米颗粒 (AuNPs) 常用于各种生物传感器。然而,其在从癌症组织中提取 DNA 方面的应用尚未得到广泛研究。从癌症组织中纯化 DNA 是诊断和治疗开发的重要步骤。几乎所有的宫颈癌病例都与高危型人乳头瘤病毒 (HR-HPV) 感染有关。到目前为止,病毒的准确诊断一直依赖于从福尔马林固定、石蜡包埋 (FFPE) 组织样本中提取足够量的 DNA。到目前为止,还没有专门针对从 FFPE 组织中提取 HR-HPV 的敏感和特异性 DNA 纯化方法。由于市售的纯化试剂盒对 HR-HPV DNA 靶标不够敏感和特异,因此本研究设计了一种基于 AuNPs 的 DNA 纯化方法,从宫颈癌组织样本中纯化足够量的 HR-HPV DNA。AuNPs 被一系列寡核苷酸探针包裹,以与 HR-HPV 基因型的特定 DNA 序列杂交。结果表明,与标准商业试剂盒 (Qiagen FFPE) 相比,733 个拷贝的 HPV 型特异性 DNA 得到了完全特异性回收,而只有 36 个拷贝得到了回收。AuNPs-探针纯化方法的主要优点是 DNA 的高产量 (91.6%)。

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