相关实验视频

Updated: Mar 21, 2026

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

Published on: April 11, 2016

没有放大或标记的DNA生物标记物少于10个副本的量化

Yoonhee Lee, Youngkyu Kim, Donggyu Lee

¹Nanogea Inc. , 6162 Bristol Parkway, Culver City, California 90230, United States.

Journal of the American Chemical Society

|May 14, 2016

概括

这项研究引入了一种新的原子力显微镜 (AFM) 检测DNA的方法. 这种技术在没有放大的情况下量化单个DNA副本,提供了一个敏感的诊断工具.

科学领域:

纳米技术和生物物理学
分子诊断

背景情况:

聚合酶链反应 (PCR) 是一种敏感的核酸检测方法,但由于目标分子数量较低而困难.
现有的检测罕见DNA生物标记的方法通常需要放大或修改,这可能引入错误或偏差.

研究的目的:

开发和验证使用基于力距离 (FD) 曲线的原子力显微镜 (AFM) 的新型量化方法.
评估在没有放大的情况下检测和量化很少数量的DNA生物标志物 (1-10副本) 的可行性.

主要方法:

使用基于FD曲线的AFM对纳米分辨率的小探测器DNA斑点 (1.4-1.9μm直径) 进行图像定位.
使用合成BCR-ABL融合基因序列作为目标DNA.
将整个探测器点映射出来,以量化检测到的探测器集群与目标副本数量相关.

主要成果:

在目标副本数量和检测到的探针集群之间实现了高相关性 (r2 = 0.994).
在单复制水平上成功检测到BCR-ABL生物标志物,尽管需要进行多次测量.
已证明适用于少于10个DNA生物标记拷贝的定量分析.

结论:

基于FD曲线的AFM成像提供了一种无标签,无放大方法来定量敏感的DNA生物标志物.
这种技术在诊断应用中对罕见的核酸标的准确检测具有前景.
该方法为特定的低副本检测场景提供了可行的PCR替代方案.

更多相关视频

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Published on: September 24, 2015

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

相关概念视频

Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

DNA Microarrays

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...