Viral Rna Amplification
Viral Rna Amplification - 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.
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.
09:44
Rapid Amplification of cDNA Ends
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2023
Source: Pablo Sanchez Bosch2, Sean Corcoran2 and Katja Brückner1,2,3 1Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research 2Department of Cell and Tissue Biology, 3Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Rapid Amplification of cDNA Ends (RACE) is a technique that allows amplification of full-length cDNA from mRNA by extending to the 3’ or 5’ end, even without prior knowledge of the sequence (Frohman et al.,...
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.
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.
FAQs
Viral RNA-dependent RNA polymerases use an existing viral RNA molecule as a template and copy it into complementary RNA strands. This copying step increases the pool of viral genetic material available during infection. Because the process occurs within infected cells, it represents a biological replication mechanism, distinct from laboratory amplification used to measure or identify viral RNA.
Cellular replication relies on viral RNA-dependent RNA polymerases to copy RNA templates into complementary strands. Diagnostic amplification instead begins by converting viral RNA into DNA through reverse transcription, then uses PCR cycles to increase selected sequences. The two approaches share template copying but differ in the molecular form amplified, the setting, and their immediate purpose.
Reverse transcription converts the RNA target into DNA, creating a form that can enter the subsequent PCR workflow. PCR then repeatedly amplifies selected DNA sequences, producing an exponential increase that makes scarce viral genetic material measurable. Separating these roles allows an RNA-originating target to be analyzed through a laboratory amplification process designed around DNA templates.
Sensitivity describes how effectively an assay detects viral genetic material when it is present, whereas specificity concerns whether the assay identifies the intended viral target rather than an unintended sequence. These properties shape confidence in results, especially when the starting material is scarce. Interpreting amplification therefore requires attention to both detectability and the possibility of false results.
A typical workflow starts with viral RNA as the genetic material of interest. Reverse transcription first converts that RNA into DNA, after which PCR cycles amplify selected sequences exponentially. The resulting increase supports measurement or detection of the target. This sequence of steps explains why the workflow can make otherwise scarce viral material measurable without relying on cellular replication.
The method supports identification and classification of viral infections, measurement of viral load, surveillance of emerging variants, and evaluation of antiviral responses. These applications use amplified genetic signals for different purposes: determining what viral material is present, estimating its amount, tracking changes among viral forms, or assessing how viral measurements change during antiviral treatment.
Amplified viral RNA provides measurable genetic information that researchers can compare across samples or time points. In surveillance, those measurements help track emerging variants and support viral classification. In antiviral studies, changes in measured viral load can contribute to evaluating responses. The results remain dependent on assay sensitivity, specificity, and awareness of possible false findings.