Preprint Communication Version 1 Preserved in Portico This version is not peer-reviewed

Major Concerns on the Identification of Bat Coronavirus Strain RaTG13 and Quality of Related Nature Paper

Version 1 : Received: 3 June 2020 / Approved: 5 June 2020 / Online: 5 June 2020 (06:17:26 CEST)

How to cite: lin, X.; Chen, S. Major Concerns on the Identification of Bat Coronavirus Strain RaTG13 and Quality of Related Nature Paper. Preprints 2020, 2020060044. https://doi.org/10.20944/preprints202006.0044.v1 lin, X.; Chen, S. Major Concerns on the Identification of Bat Coronavirus Strain RaTG13 and Quality of Related Nature Paper. Preprints 2020, 2020060044. https://doi.org/10.20944/preprints202006.0044.v1

Abstract

A recent manuscript (Zhou, P. et al. “A pneumonia outbreak associated with a new coronavirus of probable bat origin”, Nature 579, 270–273 (2020). https://doi.org/10.1038/s41586-020-2012-7) from Wuhan Institute of Virology claimed the identification of a bat coronavirus, RaTG13, which showed 96.2% genome homology with SARS-CoV-2. In this paper, we raise the puzzling observations surrounding the identification, characterization, unique genome features of this RaTG13 strain, as well as its 100% nucleotide identity in partial RdRp gene with another bat coronavirus strain BtCoV/4991. And the paper presented premature hypothesis of potential bat origin of SARS-CoV-2 while RaTG13 strain was not successfully isolated. We also present the concerns on the methodology, data quality and experiment procedures described in this paper. We call for the authors to provide additional data, to share related samples to be verified and further characterized by other scientists.

Keywords

Epidemiology; COVID-19; coronavirus; bat; RaTG13; BtCoV/4991; SARS-CoV-2; Pangolin Coronavirus; next generation sequencing

Subject

Biology and Life Sciences, Immunology and Microbiology

Comments (3)

Comment 1
Received: 14 June 2020
The commenter has declared there is no conflict of interests.
Comment: The only hint that SARS-CoV-2 is NOT genetically fabricated, but of natural origin, is this RaTG13. If RaTG13 was just a string of letters, fraudulently entered into international databases - and without any live/dead (feces) samples, this would make a completely new story. Apparently, Li and WIV are unable to deliver lab specimen on RaTG13.
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Comment 2
Received: 24 June 2020
Commenter: Jason Golias
The commenter has declared there is no conflict of interests.
Comment: Dear Authors,

I have the following hypothesis regarding RaTG13 that I feel is worthy of investigation

RaTG13 is the virus which infected 6 miners working in mine in Mojiang starting April 4th, 2012. http://eng.oversea.cnki.net/Kcms/detail/detail.aspx?filename=1013327523.nh&dbcode=CMFD&dbname=CMFD2014 (I am drafting English translation of this study from the Chinese version presently).

Shi Zhengli & research team from the Wuhan Institute of Virology arrive in June 2012 & perform 18 month surveillance campaign in Mojiang mineshaft, as cited in your paper. The reason for them coming to that mineshaft is never stated in published paper & another red flag regarding RaTG13's history.
https://europepmc.org/article/med/26920708
Isolation of RaTG13 (RaBtCoV/4991) from bats in this cave. The virus is fully sequenced at this time, & scientific community is not informed of its existence, or its direct relation to the miner's severe pneumonia. What work was performed with RaTG13 (RaBtCov/4991) in ensuing years leading up to 2019 is of utmost importance.

I propose an experiment to recreate RaTG13 infectious clone via the following methods from the published sequence:
https://www.sciencedirect.com/science/article/pii/S1931312820302316 https://www.nature.com/articles/s41586-020-2294-9

The RaTG13 infectious clone thus obtained would be used to demonstrate the virulence (similar to SARS-CoV-2) & limited airborne transmissibility of the RaTG13 virus.
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Comment 3
Received: 15 September 2020
Commenter: Stanley
The commenter has declared there is no conflict of interests.
Comment: Indeed, the partial RdRp sequence from BtCoV/4991 (370nt) is 100% identical to that of RaTG13, however, the full-length RdRp has 2795 nt, even if the 370 nt is completely identical, it does not mean the full-length RdRp sequence would be the same. This part of sequence (370nt) is highly conserved, and only 7 nt changes in SARS-CoV-2 compared to RaTG13 or BtCoV/4991. The NGS sequencing raw data of RaTG13 has been accessible online, If interested, anyone could look into it. Nevertheless, I also believe more evidence is warranted to distinguish the RdTG13 and BtCoV/4991.

The claim the WIV made that the sequence was obtained on 1/2/20, and the virus was isolated on 1/5/20 is workable in my view. As far as I know, there were at least two groups in China at the end of December, 2019 to try to identify the causative agent of novel pneumonia. As early as 12/27/19, the novel coronavirus sequence was obtained by another group. Even WIV obtained the sequence on 1/2/20, they still had enough time to get the virus isolate from cell culture. For instance, the clinical samples from patients could be inoculated with Vero cells before the viral genome from clinical samples was obtained by NGS. Once the genome was available, the primers could be designed immediately and sent out for synthesis. The primer synthesis in China will be done in less than one day, and the next day the primers could be delivered and ready to use.

I think the origin of RaTG13 and BtCoV/4991 should be addressed, and maybe the sole important point in the manuscript.
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