基因表达的比较分析 分析方法用于RNA在现场杂交 图像
Valeria Ariotta1, Eros Azzalini2, Vincenzo Canzonieri3
1Research Program in Systems Oncology, Research Programs Unit, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
The Journal of molecular diagnostics : JMD
|July 28, 2024
概括
这项研究比较了在卵巢癌中量化RNA in situ杂交 (RNA-ISH) 基因表达的方法. 像QuantISH和QuPath这样的自动化工具与RNAscope有很好的一致性,为空间基因分析提供了可访问的替代方案.
科学领域:
- 在瘤学瘤学.
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 基因表达分析在癌症研究和临床环境中至关重要.
- 传统的方法往往缺乏空间上下文,这对于理解组织微环境至关重要.
- 在位杂交RNA (RNA-ISH) 保存空间信息,使其对基因表达研究有价值.
研究的目的:
- 为了比较RNA-ISH的不同量化方法的准确性和一致性.
- 为了对RNAscope得分进行自动化工具 (QuantISH,QuPath) 和RT-droplet数字PCR的评估.
- 评估这些方法在特定基因 (CCNE1,WFDC2,PPIB) 的高度血清性卵巢癌样本中的性能.
主要方法:
- 使用了RNAscope得分,RT-droplet数字PCR,QuantISH和QuPath用于RNA-ISH量化.
- 分析了高等级的甲固定,嵌 (FFPE) 血清性卵巢癌组织.
- 专注于量化CCNE1,WFDC2和PPIB基因的表达水平.
主要成果:
- 自动化量化方法 (QuantISH,QuPath) 与RNAscope评分之间证明了良好的一致性.
- 与自动化方法相比,观察到与RT-droplet数字PCR的一致性较低.
- 量子ISH表现出强的表现,甚至有效量化了像CCNE1.1这样的低表达基因.
结论:
- 像QuantISH和QuPath这样的自动化量化工具是RNAscope用于RNA-ISH分析的可靠替代方案.
- 量子ISH的模块化设计提高了其在基因表达研究中的可访问性和实用性.
- 这些发现支持使用经过验证的自动化方法在FFPE组织中进行空间基因表达分析.
相关概念视频
02:31In-situ Hybridization
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In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
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02:34DNA Microarrays
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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...
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02:07FISH - Fluorescent In-situ Hybridization
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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
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