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10.1136/bmjresp-2020-000830

http://scihub22266oqcxt.onion/10.1136/bmjresp-2020-000830
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33627333!7907832!33627333
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suck abstract from ncbi


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pmid33627333      BMJ+Open+Respir+Res 2021 ; 8 (1): ä
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  • Assessment of SARS-CoV-2 infectivity of upper respiratory specimens from COVID-19 patients by virus isolation using VeroE6/TMPRSS2 cells #MMPMID33627333
  • Yamada S; Fukushi S; Kinoshita H; Ohnishi M; Suzuki T; Fujimoto T; Saijo M; Maeda K
  • BMJ Open Respir Res 2021[Feb]; 8 (1): ä PMID33627333show ga
  • BACKGROUND: An outbreak of novel coronavirus (SARS-CoV-2)-associated respiratory infectious diseases (COVID-19) emerged in 2019 and has spread rapidly in humans around the world. The demonstration of in vitro infectiousness of respiratory specimens is an informative surrogate for SARS-CoV-2 transmission from patients with COVID-19; accordingly, viral isolation assays in cell culture are an important aspect of laboratory diagnostics for COVID-19. METHODS: We developed a simple and rapid protocol for isolating SARS-CoV-2 from respiratory specimens using VeroE6/TMPRSS2 cells, a cell line that is highly susceptible to the virus. We also investigated a correlation between isolation of SARS-CoV-2 and viral load detected by real-time RT-PCR (rRT-PCR) using N2 primer/probe set that has been developed for testing of COVID-19 in Japan. RESULTS: The SARS-CoV-2 isolation protocol did not require blind passage of inoculated cells and yielded the results of viral isolation within 7 days after inoculation. Specimens with cycle threshold (Ct) values of <20.2, determined by rRT-PCR, were predicted to be isolation-positive. On the other hand, 6.9% of specimens with Ct values >35 were virus isolation-positive, indicating that low viral loads (high Ct values) in upper respiratory specimens do not always indicate no risk of containing transmissible virus. CONCLUSION: In combination with rRT-PCR, the SARS-CoV-2 isolation protocol provides a means for assessing the potential risk of transmissible virus in upper respiratory specimens.
  • |Animals[MESH]
  • |COVID-19 Nucleic Acid Testing[MESH]
  • |COVID-19/*transmission[MESH]
  • |Cell Line[MESH]
  • |Chlorocebus aethiops[MESH]
  • |Cytopathogenic Effect, Viral[MESH]
  • |Humans[MESH]
  • |Nasal Cavity/virology[MESH]
  • |Nasopharynx/virology[MESH]
  • |SARS-CoV-2/isolation & purification/*pathogenicity[MESH]
  • |Saliva/virology[MESH]
  • |Serine Endopeptidases/genetics[MESH]
  • |Specimen Handling[MESH]


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