Background: Evaluation of infectious virus titer is a challenge for respiratory syncytial virus (RSV) clinical trials because of the labile nature of RSV and rapid loss of infectivity without proper specimen handling. However, there has been no rigorous investigation into RSV stability in clinical specimens.
Methods: RSV stability was investigated by evaluating virus titers and defined as titer variation from baseline within three standard deviations of our titration assay. RSV stability in viral transport medium (VTM) at 4°C and the effect of freezing method on stability were evaluated using RSV-A2 stock. RSV stability in nasal aspirates collected in VTM at 4°C was estimated by regression analysis of virus titers measured at several time points. Stability of these specimens stored at -80°C for 10-15 months after freezing by the method, which maintained RSV-A2 stability, was also assessed.
Results: Three standard deviations were calculated from our titration assay as 0.97 log10 50% tissue culture infectious dose (TCID50/mL), and RSV stability was defined as variation of virus titer from baseline within 1.0 log10TCID50/mL. RSV-A2 in VTM at 4°C was stable for at least 120 h. Freezing at -80°C negatively affected virus stability, whereas freezing in liquid nitrogen or a dry ice-ethanol bath did not. RSV in nasal aspirates was stable for 2 days at 4°C and for 10-15 months at -80°C after snap freezing.
Conclusions: RSV in nasal aspirates in VTM was estimated to be stable for 2 days at 4°C and for approximately 1 year at -80°C.
Keywords: nasal aspirates; respiratory syncytial virus; virus stability; virus titer.
© 2024 The Author(s). Influenza and Other Respiratory Viruses published by John Wiley & Sons Ltd.