The DOI matches its Crossref record; 11 of the 11 references it cites were found in a registry or an index.
Viral Concentration Determination Through Plaque Assays: Using Traditional and Novel Overlay Systems
Open accessAbstract
Plaque assays remain one of the most accurate methods for the direct quantification of infectious virons and antiviral substances through the counting of discrete plaques (infectious units and cellular dead zones) in cell culture. Here we demonstrate how to perform a basic plaque assay, and how differing overlays and techniques can affect plaque formation and production. Typically solid or semisolid overlay substrates, such as agarose or carboxymethyl cellulose, have been used to restrict viral spread, preventing indiscriminate infection through the liquid growth medium. Immobilized overlays restrict cellular infection to the immediately surrounding monolayer, allowing the formation of discrete countable foci and subsequent plaque formation. To overcome the difficulties inherent in using traditional overlays, a novel liquid overlay utilizing microcrystalline cellulose and carboxymethyl cellulose sodium has been increasingly used as a replacement in the standard plaque assay. Liquid overlay plaque assays can be readily performed in either standard 6 or 12 well plate formats as per traditional techniques and require no special equipment. Due to its liquid state and subsequent ease of application and removal, microculture plate formats may alternatively be utilized as a rapid, accurate and high throughput alternative to larger scale viral titrations. Use of a non heated viscous liquid polymer offers the opportunity to streamline work, conserves reagents, incubator space, and increases operational safety when used in traditional or high containment labs as no reagent heating or glassware are required. Liquid overlays may also prove more sensitive than traditional overlays for certain heat labile viruses.
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Cites
New low-viscosity overlay medium for viral plaque assays.
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Replication and Plaque Assay of Influenza Virus in an Established Line of Canine Kidney Cells
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The Plaque Assay of Animal Viruses
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Self-citationStandardization of the Filovirus Plaque Assay for Use in Preclinical Studies
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Abreu, C. M., Gama, L., Krasemann, S., Chesnut, M., Odwin-Dacosta, S., Hogberg, H. T., Hartung, T., & Pamies, D. (2018) · Frontiers in Microbiology · doi:10.3389/fmicb.2018.02766 · Cited by 123
Manufacturing of recombinant adeno-associated viruses using mammalian expression platforms
Robert, M., Chahal, P. S., Audy, A., Kamen, A., Gilbert, R., & Gaillet, B. (2017) · Biotechnology Journal · doi:10.1002/biot.201600193 · Cited by 89
Tristan, C. A., Ormanoglu, P., Slamecka, J., Malley, C., Chu, P.-H., Jovanovic, V. M., Gedik, Y., Jethmalani, Y., Bonney, C., Barnaeva, E., et al. (2021) · Stem Cell Reports · doi:10.1016/j.stemcr.2021.11.004 · Cited by 80
Bracey, D. N., Seyler, T. M., Jinnah, A. H., Smith, T. L., Ornelles, D. A., Deora, R., Parks, G. D., Van Dyke, M. E., & Whitlock, P. W. (2019) · Xenotransplantation · doi:10.1111/xen.12534 · Cited by 25
Metadata
- Authors
- Baer, A., & Kehn-Hall, K.
- Venue
- Journal of Visualized Experiments
- Publisher
- MyJove Corporation
- Published
- 2014-11-04
- Issue
- 93
- Pages
- e52065-e52065
- ISSN
- 1940-087X (ISSN-L)
- OpenAlex
- W1970338528
- Cited by
- 441
- References
- 11
- FWCI
- 7.19
- Type
- journal article
- Language
- en
- Open access
- bronze
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