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Thanks so much, Mike. I'll post a question now.
Hello Mike,
I have some questions.
(1) Do you know of any published valid "SARS-CoV-2" negative controls for CPE (cytopathic effect)? (excluding unpublished Stefan Lanka's experiments).
I haven't found anything in the literate thus far (pubmed, google etc). They are either omitted, invalid or undocumented.
There is this paper https://www.nature.com/articles/s41467-021-22166-4 but this for cell death not CPE ("Importantly, we saw no differences in cell number between mock and SARS-CoV-2 infected cells at the time of metabolite extraction 24 h post SARS-CoV-2 infection, suggesting no measurable difference in cell death") and the full materials and methods of the mock are not documented.
I am aware of these studies but they are not for SARS-CoV-2 and are quite dated.
ENDERS JF, PEEBLES TC. Propagation in tissue cultures of cytopathogenic agents from patients with measles. Proc Soc Exp Biol Med. 1954;86(2):277-286. https://pubmed.ncbi.nlm.nih.gov/13177653/
RUSTIGIAN R, JOHNSTON P, REIHART H. Infection of monkey kidney tissue cultures with virus-like agents. Proc Soc Exp Biol Med. 1955;88(1):8-16. https://https://pubmed.ncbi.nlm.nih.gov/14357329/
COHEN SM, GORDON I, RAPP F, MACAULAY JC, BUCKLEY SM. Fluorescent antibody and complement-fixation tests of agents isolated in tissue culture from measles patients. Proc Soc Exp Biol Med. 1955;90(1):118-122. https://pubmed.ncbi.nlm.nih.gov/13273372/
BECH V, VON MAGNUS P. Studies on measles virus in monkey kidney tissue cultures. 1. Isolation of virus from 5 patients with measles. Acta Pathol Microbiol Scand. 1958;42(1):75-85. https://pubmed.ncbi.nlm.nih.gov/13508250/
Magnus, P.v., Andersen, E.K., Petersen, K.B. and Birch-Andersen, A. (1959), A POX-LIKE DISEASE IN CYNOMOLGUS MONKEYS. Acta Pathologica Microbiologica Scandinavica, 46: 156-176. https://doi.org/10.1111/j.1699-0463.1959.tb00328.x
(2) In this paper https://doi.org/10.1016/j.cell.2020.05.042 on page 431 under Figure 1(C) it states
"CPE and GFP signals were observed in Vero-E6 cells electroporated with sub-genomic RNA (sgRNA)-N alone (mock) or sgRNA-N mixed with full-length RNA transcripts (recombinant viruses) at two days after transfection. Scale bar, 100 mm."
(a) Do you know what "sub-genomic RNA (sgRNA)-N alone" means?
(b) Would you consider this as a valid negative control (i.e., no "SARS-CoV-2") for CPE?
Thank You,
Dmitry