Improved plaque assay for human coronaviruses 229E and OC43
| Autor: | Han Chi Pan, Nicole Bracci, Shih-Chao Lin, Caitlin W. Lehman, Kylene Kehn-Hall |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
viruses medicine.disease_cause General Biochemistry Genetics and Molecular Biology Virus 03 medical and health sciences Immune system Biosafety level biology.animal Virology Agarose Medicine Mink 229E 030304 developmental biology Coronavirus Virus quantification 0303 health sciences biology 030306 microbiology business.industry SARS-CoV-2 General Neuroscience virus diseases COVID-19 General Medicine OC43 Titer General Agricultural and Biological Sciences business Biotechnology Plaque assay |
| Zdroj: | PeerJ |
| ISSN: | 2167-8359 |
| Popis: | In light of the COVID-19 pandemic, studies that work to understand SARS-CoV-2 are urgently needed. In turn, the less severe human coronaviruses such as HCoV-229E and OC43 are drawing newfound attention. These less severe coronaviruses can be used as a model to facilitate our understanding of the host immune response to coronavirus infection. SARS-CoV-2 must be handled under biosafety level 3 (BSL-3) conditions. Therefore, HCoV-229E and OC43, which can be handled at BSL-2 provide an alternative to SARS-CoV-2 for preclinical screening and designing of antivirals. However, to date, there is no published effective and efficient method to titrate HCoVs other than expensive indirect immunostaining. Here we present an improved approach using an agarose-based conventional plaque assay to titrate HCoV 229E and OC43 with mink lung epithelial cells, Mv1Lu. Our results indicate that titration of HCoV 229E and OC43 with Mv1Lu is consistent and reproducible. The titers produced are also comparable to those produced using human rhabdomyosarcoma (RD) cells. More importantly, Mv1Lu cells display a higher tolerance for cell-cell contact stress, decreased temperature sensitivity, and a faster growth rate. We believe that our improved low-cost plaque assay can serve as an easy tool for researchers conducting HCoV research. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|