1. Performing the SOP with both HRV-16 and FMDV to identify where loss of genomic RNA infectivity occurs.
Performing the SOP with both HRV-16 and FMDV to identify where loss of genomic RNA infectivity occurs. (A) Flow chart depicting a test for HRV-16 or FMDV loss of RNA infectivity. Briefly, 60 tubes of HRV-16 gRNA were subject to six different conditions in replicates of 10. The SOP was performed, and a subset was purified at each step and tested for the presence of infectious RNA over three blind passages on H1 HeLa cells. For FMDV, viral RNA, intermediates, or the final SOP products were electroporated into LFBK αvβ6 cells. (B) Results of infectivity testing with HRV-16. Each symbol represents the value for an individual sample. Samples to the right of the red line highlight steps where all samples tested had no detectable infectious HRV-16 genomic RNA. The “Viral RNA +RNase” group demonstrates the RNase treatment is sufficient to inactivate all infectious gRNA. (C) Results of infectivity testing with FMDV as outlined in panel A. Each symbol represents the value for an individual sample. Samples to the right of the red line highlight steps where all samples tested had no detectable infectious FMDV genomic RNA.
Lindsey A. Moser et al. mSystems 2016; doi: /mSystems