Susceptibility to SARS-CoV-2 of Cell Lines and Substrates Commonly Used to Diagnose and Isolate Influenza and Other Viruses

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1. Version published to 10.3201/eid2705.210023

   May 1, 2021
2. 

   ScreenIT

   Jan 6, 2021

   SciScore for 10.1101/2021.01.04.425336: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cell lysates and recombinant ACE2 protein control (Sino Biological) were immunoblotted for ACE2 and β-actin using primary antibodies (1:500 polyclonal goat anti-human ACE2, R&D Systems,	ACE2 suggested: (LSBio (LifeSpan Cat# LS-C347-500, RRID:AB_1271966) anti-human ACE2 suggested: None
   1:1000 monoclonal mouse anti-β-Actin, Abcam, AB8226) followed by secondary antibodies (1:4000 donkey anti-goat, Abcam; 1:4000 goat anti-mouse, Biorad).	anti-β-Actin suggested: None a…

   SciScore for 10.1101/2021.01.04.425336: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cell lysates and recombinant ACE2 protein control (Sino Biological) were immunoblotted for ACE2 and β-actin using primary antibodies (1:500 polyclonal goat anti-human ACE2, R&D Systems,	ACE2 suggested: (LSBio (LifeSpan Cat# LS-C347-500, RRID:AB_1271966) anti-human ACE2 suggested: None
   1:1000 monoclonal mouse anti-β-Actin, Abcam, AB8226) followed by secondary antibodies (1:4000 donkey anti-goat, Abcam; 1:4000 goat anti-mouse, Biorad).	anti-β-Actin suggested: None anti-goat suggested: None anti-mouse suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   SARS-CoV-2/Massachusetts/VPT1/2020 (MA/VPT1) was isolated in Vero E6 cells from a nasopharyngeal specimen collected in April 2020.	Vero E6 suggested: None
   MDCK-hCK cells were kindly provided by Y. Kawaoka (University of Wisconsin-Madison).	MDCK-hCK suggested: None
   MDCK-NBL2, Vero E6, CV-1, A549, CRFK, Mv1Lu, RD, Hep-2c, HeLa, and L20B cells were obtained from American Type Culture Collection (ATCC) or maintained at CDC’s Division of Scientific Resources	MDCK-NBL2 suggested: CLS Cat# 602280/p823_MDCK_(NBL-2, RRID:CVCL_0422) A549 suggested: None Hep-2c suggested: ECACC Cat# 85020207, RRID:CVCL_2940) HeLa suggested: None L20B suggested: None
   All 25 cell lines listed in Table 1 were obtained from Quidel Corporation (San Diego, CA) in pre-seeded 24-well plates except for CRFK and RhMK cells, which were obtained in T-75 flasks and seeded into 24-well plates in the lab one day prior to infection.	RhMK suggested: ATCC Cat# CRL-6317, RRID:CVCL_U238)
   Exogenous expression of ACE2 in MDCK cells: Constructs co-expressing full-length hACE2 or cACE2 with mCherry2 protein (CMV promoter-ACE2-IRES-mCherry2) were generated and transfected into MDCK-SIAT1 cells via electroporation with Lonza Nucleofector system (Lonza) using the manufacturer’s protocol with program A024.	MDCK suggested: None
   1.5×106 MDCK-SIAT1 cells were transfected with 10 μg DNA (pCMV-hACE2-IRES-mCherry2, pCMV-cACE2-IRES-mCherry2, or pCMV-IRES-mCherry2 empty control).	MDCK-SIAT1 suggested: ECACC Cat# 05071502, RRID:CVCL_Z936)
   Software and Algorithms
   Sentences	Resources
   Virus infection of embryonated chicken eggs: Specific-pathogen-free (SPF) embryonated chicken eggs were obtained from Charles River Laboratories (North Franklin, CT, USA).	Charles River Laboratories suggested: (Charles River Laboratories, RRID:SCR_003792)
   ACE2 Sequence alignment: ACE2 protein sequences for human (NP_001358344.1), African green monkey (AAY57872.1), rhesus macaque (ACI04564.1), mouse (NP_001123985.1), dog (XP_005641049.1), cat (NP_001034545.1), American mink (QPL12211), and chicken (XP_416822.2) were aligned using MUSCLE alignment in Geneious Prime software (version 2019.2.3).	MUSCLE suggested: (MUSCLE, RRID:SCR_011812)

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   Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

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   Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   Results from rtransparent:

   • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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3. 

   Version published to 10.1101/2021.01.04.425336 on bioRxiv

   Jan 5, 2021