Functional and genetic analysis of viral receptor ACE2 orthologs reveals a broad potential host range of SARS-CoV-2

Abstract

COVID-19, caused by SARS-CoV-2, is a major global health threat. The host range of SARS-CoV-2 and intermediate hosts that facilitate its transmission to humans remain unknown. We found that SARS-CoV-2 has the potential to infect a broad range of mammalian hosts, including domestic animals, pets, livestock, and animals commonly found in zoos and aquaria. Those species may be at risk for human-to-animal or animal-to-animal transmissions of SARS-CoV-2. Our study highlights the importance of banning illegal wildlife trade and consumption, and enforcing the importance of surveilling animals in close contact with humans as potential zoonotic reservoirs to prevent outbreaks in the future.

SciScore for 10.1101/2020.04.22.046565: (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	Contamination: Cells were tested routinely and found to be free of mycoplasma contamination.

Table 2: Resources

Antibodies
Sentences	Resources
The blots were exposed to primary antibodies anti-β-Tubulin (CW0098, CWBIO), or anti-FLAG (F7425, Sigma) in 5% nonfat milk in 1× PBS containing 0.1% Tween 20 for 2 h.	anti-β-Tubulin ( CW0098 suggested: None anti-FLAG suggested: None
Cells were fixed with 4% paraformaldehyde in PBS, permeablized with 0.2% Triton X-100, and incubated with the rabbit polyclonal antibody against …

SciScore for 10.1101/2020.04.22.046565: (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	Contamination: Cells were tested routinely and found to be free of mycoplasma contamination.

Table 2: Resources

Antibodies
Sentences	Resources
The blots were exposed to primary antibodies anti-β-Tubulin (CW0098, CWBIO), or anti-FLAG (F7425, Sigma) in 5% nonfat milk in 1× PBS containing 0.1% Tween 20 for 2 h.	anti-β-Tubulin ( CW0098 suggested: None anti-FLAG suggested: None
Cells were fixed with 4% paraformaldehyde in PBS, permeablized with 0.2% Triton X-100, and incubated with the rabbit polyclonal antibody against SARS-CoV nucleocapsid protein (Rockland, 200-401-A50, 1μg/ml) at 4 °C overnight.	SARS-CoV nucleocapsid protein suggested: (Creative Diagnostics Cat# DMAB8869, RRID:AB_2392503)
After three washes, cells were incubated with the secondary goat anti-rabbit antibody conjugated with Alexa Fluor 488 (Thermo #A11034, 2 μg/ml) for 2 h at room temperature, followed by staining with 4’,6-diamidino-2-phenylindole (DAPI).	anti-rabbit suggested: (Thermo Fisher Scientific Cat# A-11034, RRID:AB_2576217)
Experimental Models: Cell Lines
Sentences	Resources
Cell cultures and SARS-CoV-2 virus: HEK293T cells (American Tissue Culture Collection, ATCC, Manassas, VA, CRL-3216), Vero E6 (Cell Bank of the Chinese Academy of Sciences, Shanghai, China) and HeLa (ATCC #CCL-2) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Gibco, NY, USA) supplemented with 10% (vol/vol	Vero E6 suggested: None
(Addgene #12259) and psPAX2 (Addgene #12260) and the transfer vector with VigoFect DNA transfection reagent (Vigorous) into HEK293T cells.	HEK293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
Immunofluorescence staining of viral nucleocapsids: HeLa cells were transduced with lentiviruses expressing the ACE2 from different species for 48 h.	HeLa suggested: None
Software and Algorithms
Sentences	Resources
The alignment was conducted using the MUSCLE algorithm 29.	MUSCLE suggested: (MUSCLE, RRID:SCR_011812)
Images were processed using the ImageJ program (http://rsb.info.nih.gov/ij/).	ImageJ suggested: (ImageJ, RRID:SCR_003070)

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).

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.

Results from TrialIdentifier: No clinical trial numbers were referenced.

Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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

Results from rtransparent:

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

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Functional and genetic analysis of viral receptor ACE2 orthologs reveals a broad potential host range of SARS-CoV-2

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