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Evaluation of ViroCyt® Virus Counter for Rapid Filovirus Quantitation
Open accessAbstract
Development and evaluation of medical countermeasures for diagnostics, vaccines, and therapeutics requires production of standardized, reproducible, and well characterized virus preparations. For filoviruses this includes plaque assay for quantitation of infectious virus, transmission electron microscopy (TEM) for morphology and quantitation of virus particles, and real-time reverse transcription PCR for quantitation of viral RNA (qRT-PCR). The ViroCyt® Virus Counter (VC) 2100 (ViroCyt, Boulder, CO, USA) is a flow-based instrument capable of quantifying virus particles in solution. Using a proprietary combination of fluorescent dyes that stain both nucleic acid and protein in a single 30 min step, rapid, reproducible, and cost-effective quantification of filovirus particles was demonstrated. Using a seed stock of Ebola virus variant Kikwit, the linear range of the instrument was determined to be 2.8E+06 to 1.0E+09 virus particles per mL with coefficient of variation ranging from 9.4% to 31.5% for samples tested in triplicate. VC particle counts for various filovirus stocks were within one log of TEM particle counts. A linear relationship was established between the plaque assay, qRT-PCR, and the VC. VC results significantly correlated with both plaque assay and qRT-PCR. These results demonstrated that the VC is an easy, fast, and consistent method to quantify filoviruses in stock preparations.
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FV repeatability for rAAV2 and -3 is around 5% CV but commonly 10%-30% CV in independent studies on other viruses.
“Repeatability for rAAV2 and -3 is reported around 5% CV but commonly 10%–30% CV in independent studies on other viruses.”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [65]
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Metadata
- Authors
- Rossi, C., Kearney, B., Olschner, S., Williams, P., Robinson, C., Heinrich, M., Zovanyi, A., Ingram, M., Norwood, D., & Schoepp, R.
- Affiliations
- United States Army Medical Research Institute of Infectious Diseases (US)
- Corresponding author
- Rossi, Cynthia
- Venue
- Viruses
- Publisher
- MDPI AG
- Published
- 2015-02-20
- Volume
- 7
- Issue
- 3
- Pages
- 857-872
- ISSN
- 1999-4915 (ISSN-L)
- PMC
- PMC4379551
- OpenAlex
- W2148294427
- Cited by
- 46
- References
- 19
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 3 · 2022: 5 · 2021: 5 · 2020: 4 · 2019: 5 · 2018: 4 · 2017: 8 · 2016: 4 · 2015: 6
- FWCI
- 5.70
- Type
- journal article
- Language
- en
- Open access
- gold
- dx.doi.org
- Licence
- Locations
- Viruses (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Viral Infections and Outbreaks Research
- Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
- Physical Sciences › Engineering › Biomedical Engineering › Biosensors and Analytical Detection
- Keywords
- flow cytometry, fluorescent dyes, Ebola virus, plaque assay, qRT-PCR, transmission electron microscopy, medical countermeasures
- MeSH terms
- Animals, Humans, Staining and Labeling, Time Factors, Viral Load, Ebolavirus
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