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Updated: Jun 24, 2025

Detection of Viral RNA by Fluorescence in situ Hybridization FISH

Detection of Viral RNA by Fluorescence in situ Hybridization FISH

Published on: May 5, 2012

一种用于检测SARS-CoV-2的替代方法,使用修改的光 in situ 杂交

Agnieszka Sroka-Oleksiak¹, Agnieszka Krawczyk¹, Katarzyna Talaga-Ćwiertnia¹

¹Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121, Krakow, Poland.

|June 6, 2024

概括

现场光混合化 (FISH) 提供了一种用于检测SARS-CoV-2的新方法. 这种分子测试可视化病毒,补充实时逆转录酶聚合酶链反应 (rRT-PCR) 测试,以改善微生物诊断.

科学领域:

分子生物学分子生物学
病毒学病毒学
微生物学微生物学

背景情况:

实时逆转录酶聚合酶链反应 (rRT-PCR) 是SARS-CoV-2检测的标准,但可以产生虚假阴性.
需要补充诊断工具来提高SARS-CoV-2检测准确度.

研究的目的:

开发一种高度特定的分子测试,用于检测和可视化SARS-CoV-2感染.
评估一种新的光现场杂交 (FISH) 方法在SARS-CoV-2诊断中的有效性.

主要方法:

一个通用探针和具有FLAP序列的特定寡核酸被设计用于基于FISH的可视化.
使用感染SARS-CoV-2的Vero CCL-81细胞优化了FISH测定条件.
FISH方法在290个被怀疑患有SARS-CoV-2的患者的鼻抽样上得到了验证,结果与rRT-PCR进行了比较.

主要成果:

在FISH方法中,发现了220个正面结果和70个负面结果.
rRT-PCR产生了200个阳性和90个阴性结果.
在FISH和rRT-PCR之间检测率的差异是统计学上显著的 (p=0.008).

结论:

现场光杂交 (FISH) 显示出作为SARS-CoV-2微生物诊断的补充方法的前景.
开发的FISH试验证明了检测SARS-CoV-2的高特异性.
鱼类可以帮助提高SARS-CoV-2诊断的准确性,可能减少虚假阴性.

关键词:

光在现场混合化光在现场混合化.混合化探测器探测器这就是SARS-CoV-2病毒.病毒诊断病毒诊断

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相关概念视频

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

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,...

In-situ Hybridization

In-situ Hybridization

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...

Immunofluorescence Microscopy

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.