A Quantitative Real-Time PCR Assay for Measuring Poxvirus Replication and Cell Binding.

Brian M Ward, Stephanie R Monticelli

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0citing papers in PubMed

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4 · The record

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5 · Who and what money

Authors and funding

2 authors.

Brian M WardDepartment of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY, USA. Brian_Ward@urmc.rochester.edu.

Stephanie R MonticelliViral Immunology Branch, United States Army Medical Research Institute of Infectious Disease, Frederick, MD, USA.

Funding

Protein Interactions Involved in Orthopoxvirus EnvelopmentR01AI067391 · NIAID · UNIVERSITY OF ROCHESTER · PI WARD, BRIAN M · 2007 to 2017

$3.2M

Understanding the Function of F13 as a Matrix Protein for Poxvirus Intracellular EnvelopmentR01AI168023 · NIAID · UNIVERSITY OF ROCHESTER · PI BRIAN M WARD · 2023 to 2026

$1.5M

NIAID NIH HHS R01 AI067391NIAID NIH HHS R01 AI168023

6 · The paper itself

Abstract

Quantitative real-time PCR (qPCR) is a fast and reliable method to quantify viral genomes as a surrogate to titering on monolayers of cells for measuring virus replication. Whether it be for determining the number of virions released, the total number of genomes produced during infection, or the number of virions bound to a cell, qPCR assays can be adapted to quickly enumerate total viral genomes in a broad range of experiments comparing virus replication under different conditions. In addition, qPCR offers several advantages compared to plaque assays including time, linearity over 9 logs, and scalability from tens-to-hundreds of samples, depending on the qPCR machine. Here we describe a qPCR assay for quantifying vaccinia virus' dsDNA genome that can be used to determine the total number of virions produced. Furthermore, we describe a straightforward protocol for a cell-binding assay that is sensitive enough to use with small concentrations of inoculating virions. This protocol is suitable for measuring the cell-binding ability of mutations that affect virus production and infectivity.

Indexed as

Real-Time Polymerase Chain ReactionVaccinia virusVirus ReplicationCell LineDNA, ViralGenome, ViralHumansPoxviridaeVirionDNA, ViralCell-binding assayGenome quantificationMonkeypox virusOrthopoxvirusQuantitative PCRVaccinia virusVirion quantification

Identifiers

PMID39621259

DOI10.1128/jvi.71.5.4032-4041.1997

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.