Use of Flow Cytometry for Rapid, Quantitative Detection of Poliovirus-Infected Cells via TAT Peptide-Delivered Molecular Beacons

Sivaraman, Divya; Yeh, Hsiao-Yun; Mulchandani, Ashok; Yates, Marylynn V.; Chen, Wilfred

doi:10.1128/aem.02429-12

Cited by 9 publications

(5 citation statements)

References 21 publications

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“…Fluorescent labeling has been used to reliably detect and quantify viruses like HSV, CMV, adenovirus, influenza A, RSV, rubella, coronavirus, dengue, and parainfluenza [ 182 , 183 ]. In many cases, non-specific nucleic acid and protein-binding fluorescent dyes are used to label virus particles, but some reports also describe the use of specific antibodies to detect HIV-1, poliovirus, dengue virus [ 184 , 185 , 186 ]. These fluorescent labeling methods may affect particle infectivity, which is of minor concern if virometry is used only to detect and measure virus particles.…”

Section: Beyond Traditional Fluorescence Microscopymentioning

confidence: 99%

Fluorescent Protein Approaches in Alpha Herpesvirus Research

Hogue

Bosse

Engel

et al. 2015

Viruses

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In the nearly two decades since the popularization of green fluorescent protein (GFP), fluorescent protein-based methodologies have revolutionized molecular and cell biology, allowing us to literally see biological processes as never before. Naturally, this revolution has extended to virology in general, and to the study of alpha herpesviruses in particular. In this review, we provide a compendium of reported fluorescent protein fusions to herpes simplex virus 1 (HSV-1) and pseudorabies virus (PRV) structural proteins, discuss the underappreciated challenges of fluorescent protein-based approaches in the context of a replicating virus, and describe general strategies and best practices for creating new fluorescent fusions. We compare fluorescent protein methods to alternative approaches, and review two instructive examples of the caveats associated with fluorescent protein fusions, including describing several improved fluorescent capsid fusions in PRV. Finally, we present our future perspectives on the types of powerful experiments these tools now offer.

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Section: Beyond Traditional Fluorescence Microscopymentioning

confidence: 99%

Fluorescent Protein Approaches in Alpha Herpesvirus Research

Hogue

Bosse

Engel

et al. 2015

Viruses

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“…Diverse approaches have been used to identify and describe viruses in various organisms including transmission electron microscopy (TEM) as described in Tables 2-4 [129,213], PCR-based analyses [214], DNA in situ hybridization [215], immune-histochemistry [216], flow cytometry [217], next generation sequencing (NGS) [218] and metagenomic analyses [138,142].…”

Section: Detection Methods For Marine Viruses In Coral Ecosystemmentioning

confidence: 99%

A Review of Marine Viruses in Coral Ecosystem

Ambalavanan

Iehata

Fletcher

et al. 2021

JMSE

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Coral reefs are among the most biodiverse biological systems on earth. Corals are classified as marine invertebrates and filter the surrounding food and other particles in seawater, including pathogens such as viruses. Viruses act as both pathogen and symbiont for metazoans. Marine viruses that are abundant in the ocean are mostly single-, double stranded DNA and single-, double stranded RNA viruses. These discoveries were made via advanced identification methods which have detected their presence in coral reef ecosystems including PCR analyses, metagenomic analyses, transcriptomic analyses and electron microscopy. This review discusses the discovery of viruses in the marine environment and their hosts, viral diversity in corals, presence of virus in corallivorous fish communities in reef ecosystems, detection methods, and occurrence of marine viral communities in marine sponges.

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“…Numerous approaches have been used to describe viruses in various organisms including: electron microscopy (Davy and Patten, 2007;, PCR-based representational difference analysis (Chang et al, 1994), DNA in situ hybridization (Teifke et al, 2000), immuno-histochemistry (Gouda et al, 2010), flow cytometry (Sivaraman et al, 2013), and PCR and next generation sequencing (NGS) (Marston et al, 2013).…”

Section: Methods Used To Identify Viruses In Coralsmentioning

confidence: 99%

The role of viruses in coral health and disease

Sweet

Bythell

2017

Journal of Invertebrate Pathology

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Metagenomic and electron microscopy studies confirm that the coral microbiome contains a rich diversity and abundance of viruses. While there have been no definitive tests of disease causation by viruses in corals, viruses have been implicated as coral pathogens in a number of studies. Growing evidence also indicates that latent viral infections can compromise the algal symbionts under environmental stress and may be involved in the coral bleaching response. Conversely, bacteriophages and archaeal phage viruses are abundant in the microbiome of healthy corals and are likely to be involved in complex ecological networks, genetic material transfer and selective co-evolution within the surface mucus layers and tissues. The relative importance of viral control of bacterial and archaeal populations is unknown, but they are almost certain to be exerting some level of control on the composition and maintenance of the coral microbiome. While rapid leaps in the capability to detect viruses have been made due to advances in metagenomics and bioinformatics, these approaches need now to be integrated with in vitro culture and challenge experiments to assess the functional roles of viruses in health and disease, and it is imperative that interactions with other members of the coral microbiome are taken into account when assessing disease causation.

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Use of Flow Cytometry for Rapid, Quantitative Detection of Poliovirus-Infected Cells via TAT Peptide-Delivered Molecular Beacons

Cited by 9 publications

References 21 publications

Fluorescent Protein Approaches in Alpha Herpesvirus Research

Fluorescent Protein Approaches in Alpha Herpesvirus Research

A Review of Marine Viruses in Coral Ecosystem

The role of viruses in coral health and disease

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