🚨🚨🚨 Gear up. Revealing three theses from Wuhan Institute of Virology which provides new details on Mojiang sampling, 4991 genome, 7896-clade, unpublished CoVs, coronavirus reverse genetics and more! All theses were supervised by Shi Zhengli. Links at the end of the thread.
1/ Serological Cross-Reactivity Analysis of Coronavirus Based on Nucleocapsid Proteins (by Ning Wang, May 2014) ImageImage
Here are some key takeaways:

• It is the 1st time a WIV study mentions the miners outbreak: "Three miners died from pneumonia in Mojiang..we investigated the virus carried by bats in this cave..it is likely that the six miners were infected with the pathogen carried by bats."
• This very obviously - and for the third time - flies in the face of WIV's claim that the miner’s illness were thought to be related to fungal infection.
• What I find most fascinating: In this study, "fever patient sera were obtained from a hospital in Yunnan Province." 30 sera were obtained in total, and all marked MJ (almost certainly Mojiang).

Note: Shi's @Nature addendum mentions only 13 sera collected from 4 patients. Image
• A total of 170 bat samples were collected from Mojiang in 2012, of which 109 were positive for coronavirus, with a positive rate of 64%!

Seems the mine was indeed a holy grail for virus hunters!
• The author notes: "The full-length sequences of N and RdRp genes of several coronaviruses have been successfully amplified, but only a small number of full-length sequences of S genes have been amplified."
• On S gene sequences, the author notes: “there are still some urgent technical barriers to the amplification of the S gene sequences, especially the full-length sequences, and it is urgent to accomplish the relevant technical breakthroughs in future studies”
• In the acknowledgement section, the author thanks Linfa Wang for his "guidance" on the project. This confirms my long held suspicion that Linfa Wang knew about the miners outbreak.

"If three people died and it was controlled would we know it?"
• And finally the study reports, for the first time ever, Ra4991: "Overlap PCR was performed on the amplified bat coronavirus N gene, of which 4991, 3740(β)-N was amplified by laboratory associate" ImageImage
2/ Reverse Genetic System of Bat SARS-like Coronaviruses and Function of ORFX (by Lei-Ping Zeng, April 2017) ImageImageImage
A few illustrative quotes from the thesis:

• “In this study, we combined the advantages of two mainstream coronavirus reverse genetics methods, established a new efficient and cost-effective method”, and "a scheme to replace the S gene without traces”.
• The reverse genetics system established in this study “can be used to rescue viruses whose sequences have only been identified, to study the function of virus-encoded proteins, to help assess their pathogenicity, and to explore their evolutionary and transmission mechanisms.”
• The author also points out: “It can also help to evaluate the effectiveness of existing antibodies and drugs against SARS against these bat-derived SL-CoV.”
• And: “determining in the lab whether recombination occurs after co-infection with different SL-CoV strains and what new strains may arise..is beneficial for studying the evolutionary pattern of SL-CoV and providing a basis for preventing possible future outbreaks..” Image
• The author summarizes the outlook as: “At this point, our laboratory can establish a complete system from viral pathogen discovery, pathogen rescue and pathogen pathogenicity assessment to deepen and accelerate the epidemiological study of emerging viruses.”
• Zeng also mentions new methods that have emerged recently: “yeast recombination or in vitro recombination”, which “applied to the reverse genetics of coronaviruses could develop more efficient and economical method” for virology research. What happened to this line of study?
3/ Geographic Evolution of Bat SARS-related Coronaviruses (by Yu Ping, June 2019) ImageImageImage
• “This study conducted large-scale detection and analysis of SARS-like coronavirus in bat samples collected from twenty regions in China from 2011 to 2016." A total of 9261 samples were collected (2815 from Yunnan), and 170 SARS-like coronavirus were detected (92 from Yunnan). Image
• "In this study, the full-length RdRp sequence (2757 bp) was amplified for phylogenetic analysis of the virus, and finally 60 full-length RdRp sequences of SARS-like coronaviruses were obtained from positive samples."
• This is important: “SL-CoVs without deletions in the RBM region are able to utilize ACE2 receptors.. whereas SL-COVs with deletions in this region are not able to utilize ACE2 receptors.. Currently, bat SL-CoVs without deletions in the RBM region are only found in Yunnan.”
• Based on the phylogenetic analysis of the coronaviruses, the author concludes: "bat SARS-like coronaviruses tend to cluster together more by geographic location than by host species, suggesting that space plays a greater barrier role."
• In this study they sequenced the whole genome of Ra4991, and three other SARS-like coronaviruses. (The table could hold important clues.) Image
• And this is big: The study confirms, for the first time, that the 7896-clade from the mojiang mine were sequenced way before the covid outbreak. And more than just the RdRp fragment: i.e. the Spike of 7896; ORF8 of 7896, 7909 & 7952; and the RBD+NTD of 7905, 7909, 7924 & 7931. ImageImageImageImage
• So just to be clear: WIV had all these sequences at the time most closely related to SARS-CoV-2 and they chose not to disclose it in Jan 2020, and even after 15 months.
And that's a wrap! Here is the link for the theses and their translations:

drive.google.com/drive/folders/…

Copy-edited and translated by: @francsicodeasis (thank you so much!) and me, with generous help from Google, Adobe and DeepL.

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More from @TheSeeker268

Apr 17
One of the deeply under appreciated aspects of #OriginofCovid is that, according to EcoHealth docs, in 2018, WIV had over 180 viral strains that could bridge the gap between SARS2 and RaTG13/BANAL. Over 125 viral strains in the spike range of SARS2 (and could evade mAb/vaccines).
And thousands of samples from where the nearest relatives were found.

Add to this, the extensive US and Chinese state-funded projects in the 2018-19 timeframe, with the same kind of work (with live viruses in BSL-2 & -3) that could’ve led to SARS2.
Having some elementary knowledge of statistical probability, I would go so far as to say that while not impossible that some wild host brought it to Wuhan, the odds are like hitting a cosmic lottery.
Read 5 tweets
Apr 9
So a number of other people have been saying the same thing—that they learnt of the Wuhan outbreak in early/mid-December 2019. See some examples below.
JP Prasad, who runs Alberta's supply procurement system "heard disturbing news about a 'strange flu' in Wuhan, in early December", and began stocking up on masks and equipments.
edmontonjournal.com/opinion/column…
“Around December 20, 2019, I learned about the emergence of the coronavirus,” says a frontline doctor from a hospital in Xiaogan City, 66 kilometers away from Wuhan.
archive.ph/MnZrn
Read 8 tweets
Mar 18
#OriginOfCovid: Summary of what is known thus far..

- The precursor of SARS2 likely originated in bats in Yunnan/SE Asia.
- There is a direct and documented pathway from the regions where bats harbour these viruses to Wuhan, via WIV. Other ways of getting to Wuhan are possible.
- Wuhan Institute of Virology was the closest place where closely related viruses existed.
- We don’t know where the first cases occurred but it was first identified at the Huanan market after it came to the attention of the doctors in Wuhan in late December 2019.
- We don’t know when the outbreak really started but most studies indicate it was sometime between September and November 2019.
- Susceptible species were being sold at the Huanan market where some of the first (known) cases were identified.
Read 16 tweets
Mar 11
Daszak examines his own coronavirus research in Wuhan, and issues a clean bill of health for himself. You’ll just have to read it…
theintercept.com/2022/03/11/cov…
Daszak: humanized mice experiments weren't conducted by us.

So EcoHealth helped WIV import ans successfully breed humanized mice in 2018 and we are supposed to believe that they didn’t do that work although it was already funded by NIH? ImageImage
Read 9 tweets
Mar 10
A key question about COVID's origin has been: how it got to Wuhan? At this stage, nobody knows for sure, but let's explore the two competing scenarios:

1) SARS2 was brought to the Wuhan market.
2) SARS2 was brought to the Wuhan lab.

This is important, so follow me... /🧵
Scenario #1: SARS2 was brought to the Wuhan market

Assuming this to be true, then there were two separate spillovers. First, it spilled over from bats to an intermediate host. Next, it spilled over from the intermediate host to humans.
What we know for sure is that all genetically close relatives of SARS2 were found in bats in Yunnan or Southeast Asia – far from Wuhan. So, one would expect that the first spillover (bat to intermediate host) took place in this region, where bats harbour these viruses.
Read 36 tweets
Feb 27
Building on their previous papers, authors here present a new statistical analysis — as bulletproof science.

The data is good and layered, but the conclusions are based on indirect and questionable arguments. It is unpersuasive to me. Let me explain the reasons I say this. 🧵
The authors’ basic argument is: most of the (known) cases from Dec clustered around the market. There were raccoon dogs at the market (western section). There were +ve environment samples in that section. Therefore the virus jumped at the market. I don’t fundamentally buy that.
I will admit that the supporting data is rigorous and many points are valid, but the line of reasoning doesn’t make sense to me. So I’m absolutely not convinced by it from a logical point of view (also due to the inadequacy of the data and cherry picking facts). Follow me
Read 20 tweets

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