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新型快速廉价的临床样本 BRCA1 基因突变预测检测方法。

New, fast and cheap prediction tests for BRCA1 gene mutations identification in clinical samples.

机构信息

Faculty of Chemistry, University of Warsaw, Pasteura 1 Str., 02-093, Warsaw, Poland.

Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12 Str., 80-233, Gdańsk, Poland.

出版信息

Sci Rep. 2023 May 5;13(1):7316. doi: 10.1038/s41598-023-34588-9.

Abstract

Despite significant progress in cancer therapy, cancer is still the second cause of mortality in the world. The necessity to make quick therapeutic decisions forces the development of procedures allowing to obtain a reliable result in a quick and unambiguous manner. Currently, detecting predictive mutations, including BRCA1, is the basis for effectively treating advanced breast cancer. Here, we present new insight on gene mutation detection. We propose a cheap BRCA1 mutation detection tests based on the surface plasmon resonance (SPR) or quartz crystal microbalance with energy dissipation (QCM-D) response changes recorded during a hybridization process of an oligonucleotide molecular probe with DNA fragments, with and without the BRCA1 mutation. The changes in the morphology of the formed DNA layer caused by the presence of the mutation were confirmed by atomic force microscopy. The unique property of the developed SPR and QCM tests is really short time of analysis: ca. 6 min for SPR and ca. 25 min for QCM. The proposed tests have been verified on 22 different DNA extracted from blood leukocytes collected from cancer patients: 17 samples from patients with various BRCA1 gene mutation variants including deletion, insertion and missense single-nucleotide and 5 samples from patients without any BRCA1 mutation. Our test is a response to the need of medical diagnostics for a quick, unambiguous test to identify mutations of the BRCA1 gene, including missense single-nucleotide (SNPs).

摘要

尽管癌症治疗取得了重大进展,但癌症仍然是世界上第二大致死原因。快速做出治疗决策的必要性促使开发了能够快速、明确地获得可靠结果的程序。目前,检测预测性突变,包括 BRCA1,是有效治疗晚期乳腺癌的基础。在这里,我们提出了基因突变检测的新见解。我们提出了一种基于表面等离子体共振(SPR)或石英晶体微天平与能量耗散(QCM-D)响应变化的廉价 BRCA1 突变检测测试,该测试记录了寡核苷酸分子探针与 DNA 片段杂交过程中的响应变化,其中包括和不包括 BRCA1 突变。通过原子力显微镜证实了由突变引起的形成的 DNA 层的形态变化。开发的 SPR 和 QCM 测试的独特性质是分析时间非常短:SPR 约 6 分钟,QCM 约 25 分钟。该测试已在从癌症患者采集的血液白细胞中提取的 22 个不同的 DNA 上进行了验证:17 个样本来自具有各种 BRCA1 基因突变变体的患者,包括缺失、插入和错义单核苷酸,5 个样本来自没有任何 BRCA1 突变的患者。我们的测试是对医疗诊断快速、明确地识别 BRCA1 基因突变(包括错义单核苷酸(SNPs))的需求的回应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4f80/10163215/917a3e6e8642/41598_2023_34588_Fig1_HTML.jpg

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