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Titration of Human Coronaviruses Using an Immunoperoxidase Assay
Published on: April 28, 2008
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Improved plaque assay for human coronaviruses 229E and OC43
Nicole Bracci1, Han-Chi Pan2, Caitlin Lehman1
1Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
Peerj
|January 4, 2021
Summary
Researchers developed a low-cost plaque assay for titrating human coronaviruses (HCoVs) like 229E and OC43. This method uses mink lung epithelial cells (Mv1Lu) and offers a reproducible alternative to expensive techniques for HCoV research.
Area of Science:
- Virology
- Cell Biology
- Infectious Diseases
Background:
- The COVID-19 pandemic highlights the need for understanding SARS-CoV-2 and related human coronaviruses (HCoVs).
- Less severe HCoVs (e.g., HCoV-229E, OC43) serve as valuable models for studying coronavirus infections and host immune responses.
- Handling SARS-CoV-2 requires Biosafety Level 3 (BSL-3), while HCoV-229E and OC43 can be handled at BSL-2, making them suitable for preclinical screening and antiviral development.
Purpose of the Study:
- To develop an effective and efficient method for titrating HCoV-229E and OC43.
- To establish a cost-effective alternative to indirect immunostaining for HCoV titration.
- To evaluate the suitability of mink lung epithelial cells (Mv1Lu) for HCoV plaque assays.
Main Methods:
- An improved agarose-based conventional plaque assay was developed.
- The assay utilized mink lung epithelial cells (Mv1Lu) for titration of HCoV-229E and HCoV-OC43.
- Titers obtained with Mv1Lu cells were compared to those obtained with human rhabdomyosarcoma (RD) cells.
Main Results:
- The plaque assay using Mv1Lu cells provided consistent and reproducible titration results for HCoV-229E and OC43.
- Titers generated with Mv1Lu cells were comparable to those obtained using RD cells.
- Mv1Lu cells demonstrated superior characteristics, including higher tolerance to cell-cell contact stress, reduced temperature sensitivity, and faster growth rates.
Conclusions:
- The developed low-cost plaque assay is a viable and efficient method for titrating HCoV-229E and OC43.
- Mink lung epithelial cells (Mv1Lu) are a suitable cell line for HCoV plaque assays, offering advantages over traditional cell lines.
- This improved assay can serve as an accessible tool for researchers in the field of HCoV research.
