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一种基于纳米技术的利用流式细胞术捕获和检测核酸方法的开发。

Development of a nanotechnology-based approach for capturing and detecting nucleic acids by using flow cytometry.

作者信息

Robles-Remacho Agustín, Luque-González M Angélica, González-Casín Roberto A, Cano-Cortés M Victoria, Lopez-Delgado F Javier, Guardia-Monteagudo Juan J, Antonio Fara Mario, Sánchez-Martín Rosario M, Díaz-Mochón Juan José

机构信息

GENYO. Centre for Genomics and Oncological Research: Pfizer / University of Granada / Andalusian Regional Government, PTS Granada, Avenida de La Ilustracion, 114, 18016, Granada, Spain; Department of Medicinal and Organic Chemistry, School of Pharmacy, University of Granada, Campus Cartuja S/n, 18071, Granada, Spain; Biosanitary Research Institute of Granada (ibs.GRANADA), University Hospital of Granada/University of Granada, Avenida Del Conocimiento, S/n, 18016, Granada, Spain.

GENYO. Centre for Genomics and Oncological Research: Pfizer / University of Granada / Andalusian Regional Government, PTS Granada, Avenida de La Ilustracion, 114, 18016, Granada, Spain.

出版信息

Talanta. 2021 May 1;226:122092. doi: 10.1016/j.talanta.2021.122092. Epub 2021 Jan 9.

Abstract

Nucleic acid-based molecular diagnosis has gained special importance for the detection and early diagnosis of genetic diseases as well as for the control of infectious disease outbreaks. The development of systems that allow for the detection and analysis of nucleic acids in a low-cost and easy-to-use way is of great importance. In this context, we present a combination of a nanotechnology-based approach with the already validated dynamic chemical labeling (DCL) technology, capable of reading nucleic acids with single-base resolution. This system allows for the detection of biotinylated molecular products followed by simple detection using a standard flow cytometer, a widely used platform in clinical and molecular laboratories, and therefore, is easy to implement. This proof-of-concept assay has been developed to detect mutations in KRAS codon 12, as these mutations are highly important in cancer development and cancer treatments.

摘要

基于核酸的分子诊断对于遗传疾病的检测和早期诊断以及传染病爆发的控制具有特别重要的意义。开发能够以低成本且易于使用的方式检测和分析核酸的系统至关重要。在此背景下,我们提出了一种基于纳米技术的方法与已经经过验证的动态化学标记(DCL)技术的组合,该组合能够以单碱基分辨率读取核酸。该系统允许检测生物素化的分子产物,随后使用标准流式细胞仪进行简单检测,流式细胞仪是临床和分子实验室中广泛使用的平台,因此易于实施。此概念验证检测方法已被开发用于检测KRAS密码子12中的突变,因为这些突变在癌症发展和癌症治疗中非常重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec70/7794053/3462135497d0/fx1_lrg.jpg

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