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A hydroxypropyl methylcellulose plaque assay for human respiratory syncytial virus.
Takumi-Tanimukai, Yuka; Yamamoto, Soh; Ogasawara, Noriko; Nakabayashi, Sayaka; Mizuta, Katsumi; Yamamoto, Keisuke; Miyata, Ryo; Kakuki, Takuya; Jitsukawa, Sumito; Sato, Toyotaka; Tsutsumi, Hiroyuki; Kojima, Takashi; Takano, Kenichi; Yokota, Shin-Ichi.
Afiliação
  • Takumi-Tanimukai Y; Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan; Department of Otolaryngology-Head and Neck Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Yamamoto S; Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Ogasawara N; Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan; Department of Otolaryngology-Head and Neck Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan. Electronic address: ogasawara.n@sapmed.ac.jp.
  • Nakabayashi S; Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Mizuta K; Department of Microbiology, Yamagata Prefectural Institute of Public Health, Yamagata, Japan.
  • Yamamoto K; Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan; Department of Otolaryngology-Head and Neck Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Miyata R; Department of Otolaryngology-Head and Neck Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Kakuki T; Department of Otolaryngology-Head and Neck Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Jitsukawa S; Department of Otolaryngology-Head and Neck Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Sato T; Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan; Laboratory of Veterinary Hygiene, School/Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan; Graduate School of Infectious Diseases, Hokkaido University, Sapporo, Japan.
  • Tsutsumi H; Midorinosato, Saiseikai Otaru Hospital, Otaru, Japan.
  • Kojima T; Department of Cell Science, Research Institute for Frontier Medicine, Sapporo Medical University, Sapporo, Japan.
  • Takano K; Department of Otolaryngology-Head and Neck Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan.
  • Yokota SI; Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan.
J Virol Methods ; 304: 114528, 2022 06.
Article em En | MEDLINE | ID: mdl-35358640
ABSTRACT
Quantifying proliferative virus particles is one of the most important experimental procedures in virology. Compared with classical overlay materials, newly developed cellulose derivatives enable a plaque-forming assay to produce countable clear plaques easily. HEp-2 cells are widely used in plaque assays for human respiratory syncytial virus (RSV). It is crucial to use an overlay material to keep HEp-2 cell proliferation and prevent RSV particles from spreading over the fluid. Among four cellulose derivatives, carboxymethyl cellulose sodium salt (CMC), hydroxypropyl methylcellulose (HPMC), microcrystalline cellulose (MCC), and hydroxyethyl cellulose (HEC), we found that HPMC was the optimal overlay material because HPMC maintained HEp-2 cell proliferation and RSV infectivity. Although MCC was unsuitable for RSV, it assisted the plaque-forming by human metapneumovirus in TMPRSS2-expressing cells. Therefore, depending on the cells and viruses, it is necessary to use different overlay materials at varying concentrations.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Vírus Sincicial Respiratório Humano / Infecções por Vírus Respiratório Sincicial / Metapneumovirus Limite: Humans Idioma: En Revista: J Virol Methods Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Japão

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Vírus Sincicial Respiratório Humano / Infecções por Vírus Respiratório Sincicial / Metapneumovirus Limite: Humans Idioma: En Revista: J Virol Methods Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Japão