Propagation and Quantification of SARS-CoV-2

Area of Science:

• Virology
• Cell Biology
• Infectious Diseases

Background:

• The emergence of SARS-CoV-2 in late 2019 led to a global pandemic with significant mortality.
• Understanding SARS-CoV-2 and COVID-19 requires reliable methods for virus propagation and quantification.
• Standardized protocols are essential for facilitating research efforts worldwide.

Purpose of the Study:

• To describe standardized protocols for the propagation of SARS-CoV-2.
• To provide methods for quantifying infectious SARS-CoV-2.
• To establish a foundation for laboratories studying SARS-CoV-2.

Main Methods:

• SARS-CoV-2 propagation in Vero E6, Caco-2, and Calu-3 cell lines.
• Quantification of infectious SARS-CoV-2 using plaque assays.
• Quantification of infectious SARS-CoV-2 using immunofocus forming assays in Vero E6 cells.

Main Results:

• Established protocols for propagating SARS-CoV-2 in multiple cell types.
• Validated plaque and immunofocus forming assays for SARS-CoV-2 quantification.
• Demonstrated the utility of liquid overlays in quantification assays.

Conclusions:

• The described protocols enable the effective propagation and quantification of SARS-CoV-2.
• These methods support laboratories in developing the capacity to study SARS-CoV-2.
• The protocols contribute to ongoing research addressing the COVID-19 pandemic.