Viral Rna Detection
Viral Rna Detection - Related Videos
10:16
Detection of Viral RNA by Fluorescence in situ Hybridization (FISH)
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Cited by 35 •
2012
A fluorescence in situ hybridization (FISH) method was developed to visually detect viral genomic RNA using fluorescence microscopy. A probe is made with specificity to the viral RNA that can then be identified using a combination of hybridization and immunofluorescence techniques. This technique offers the advantage of identifying the localization of the viral RNA or DNA at steady-state, providing information on the control of intracellular virus trafficking events.
13:58
Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays
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Cited by 10 •
2011
Quantifying levels of HIV-1 RNA in plasma and sequencing single HIV-1 genomes from individuals with viral loads below the limit of detection (50-75 copies/ml) is difficult. Here we describe how to extract and quantify plasma viral RNA using a real time PCR assay that reliably measures HIV-1 RNA down to 0.3 copies/ml and how to amplify viral genomes by single genome sequencing, from samples with very low viral loads.
02:37
Detection of Zika Viral dsRNA Replication Intermediates Using Immunocytochemistry
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2026
Source: Contreras, D., et al. Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons. J. Vis. Exp. (2016)The video demonstrates the detection of Zika viral dsRNA intermediates in infected epithelial cells using immunocytochemistry. Infected cells are fixed and then blocked with a blocking solution. The primary and secondary antibodies are added. The nuclei are stained with a fluorescent dye. The labeled viral dsRNA is then seen under a fluorescence...
09:35
Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis
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Cited by 15 •
2013
This method describes the use of click chemistry to measure changes in host cell transcription after infection with the Rift Valley fever virus (RVFV) strain MP-12. Results can be visualized qualitatively via fluorescence microscopy or obtained quantitatively through flow cytometry. This method is adaptable for use with other viruses.
02:59
Extraction of Viral RNA from Surface-Bound Virions Using a Silica Spin Column
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2026
Source: Klaus, J. P. et al. Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment. J. Vis. Exp. (2016)This video demonstrates the extraction of viral RNA from surface-bound virions on epithelial cells using silica spin columns. The method includes chemical lysis, RNA stabilization, and membrane-based purification for downstream quantification of virus-cell attachment.
FAQs
Reverse transcriptase performs the essential conversion from viral RNA into complementary DNA, or cDNA. This step makes the viral genetic material compatible with amplification and measurement by reverse transcription quantitative PCR. The resulting cDNA provides a stable template for subsequent analysis, allowing fluorescence-based measurements to reveal viral material that may be present at low levels in a sample.
Sequence-specific probes bind selected viral genetic sequences rather than detecting nucleic acid indiscriminately. Their targeting helps connect a measured signal to a particular viral sequence and can support discrimination among closely related strains. This makes probe-based analysis useful when researchers need more information than simple detection, especially during targeted sequence analysis of viral material.
Both approaches can begin with reverse transcription when the target is viral RNA, but they use different amplification strategies. Reverse transcription quantitative PCR amplifies cDNA while measuring fluorescence, whereas isothermal amplification uses an alternative amplification format. The overview identifies both as molecular options, so the appropriate choice depends on the assay design and the information required.
Strain discrimination depends on analyzing targeted sequence differences within viral genetic material. Assays can use selected sequences or sequence-specific probes to associate the detected signal with a particular strain or closely related variant. This targeted approach supports more precise interpretation than detecting viral material alone, making it relevant to surveillance and investigations of viral evolution.
A typical workflow begins with viral RNA in a clinical, environmental, or research sample. Reverse transcriptase converts that RNA into complementary DNA, after which the cDNA is amplified. Fluorescence is measured during the amplification process, producing a detectable signal that indicates viral material and supports quantitative interpretation within the assay's targeted sequence design.
The approach can be applied to clinical, environmental, and research samples. In each setting, the assay examines available material for viral RNA and can detect low levels that may otherwise be difficult to identify. The resulting information supports different goals, including identifying infection, examining viral presence outside clinical settings, or studying viral biology under research conditions.
In public-health contexts, viral RNA detection contributes to disease surveillance, outbreak response, and treatment monitoring. In research, it helps investigate viral replication and evolution by revealing viral material and enabling targeted sequence analysis. These uses extend beyond a single diagnostic result, connecting molecular measurements with changes in viral presence, relatedness, and behavior over time.