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Improved plaque assay for human coronaviruses 229E and OC43.
Bracci, Nicole; Pan, Han-Chi; Lehman, Caitlin; Kehn-Hall, Kylene; Lin, Shih-Chao.
Afiliação
  • Bracci N; Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
  • Pan HC; National Laboratory Animal Center, National Applied Research Laboratories, Taipei, Taiwan.
  • Lehman C; Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
  • Kehn-Hall K; Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, USA.
  • Lin SC; Bachelor Degree Program in Marine Biotechnology, National Taiwan Ocean University, Keelung, Taiwan.
PeerJ ; 8: e10639, 2020.
Article em En | MEDLINE | ID: mdl-33391888
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.
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Texto completo: 1 Base de dados: MEDLINE Idioma: En Revista: PeerJ Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Base de dados: MEDLINE Idioma: En Revista: PeerJ Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Estados Unidos