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Updated: Jul 5, 2026
10:16
Detection of Viral RNA by Fluorescence in situ Hybridization (FISH)
Published on: May 5, 2012
In situ hybridization for detection of HIV RNA
1Molecular Histology, Gaithersburg, Maryland, USA.
Current Protocols in Immunology
|April 25, 2008
Summary
In situ hybridization offers a sensitive method for detecting HIV expression by identifying virion RNA. This technique involves preparing probes, hybridizing them to cells, and analyzing results using advanced microscopy.
Area of Science:
- Molecular Biology
- Virology
- Immunology
Background:
- In situ hybridization (ISH) is a powerful technique for visualizing nucleic acids within cells and tissues.
- ISH has been applied to Human Immunodeficiency Virus (HIV) research for detecting viral components like RNA and DNA.
Purpose of the Study:
- To detail protocols for using in situ hybridization to detect HIV expression, focusing on virion RNA identification.
- To establish ISH as a sensitive method for detecting HIV expression.
Main Methods:
- Preparation of radioactive or nonradioactive RNA probes.
- Hybridization of probes to cells and tissue sections.
- Detection using emulsion autoradiography, slide processing, and various microscopy techniques (bright-field, dark-field, confocal).
Main Results:
- In situ hybridization enables the identification of virion RNA, mRNA, and proviral DNA.
- Detection of virion RNA via ISH is presented as the most sensitive method for assessing HIV expression.
Conclusions:
- In situ hybridization protocols provide a comprehensive approach for detecting HIV RNA in biological samples.
- This methodology supports sensitive and detailed analysis of HIV expression at the cellular level.
Related Concept Videos
02:31In-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...
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...
02:07FISH - 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,...