https://t.co/hXlo8qgkD0
Look like that they got a classical case of PCR Cross-Contamination.
They had 2 fabricated samples (SRX9714436 and SRX9714921) on the same PCR run. Alongside with Lung07. They did not perform metagenomic sequencing on the “feces” and they did not get

A positive oral or anal swab from anywhere in their sampling. Feces came from anus and if these were positive the anal swabs must also be positive. Clearly it got there after the NA have been extracted and were from the very low-level degraded RNA which were mutagenized from
The Taq. https://t.co/yKXCgiT29w to see SRX9714921 and SRX9714436.
Human+Mouse in the positive SRA, human in both of them. Seeing human+mouse in identical proportions across 3 different sequencers (PRJNA573298, A22, SEX9714436) are pretty straight indication that the originals
Were already contaminated with Human and mouse from the very beginning, and that this contamination is due to dishonesty in the sample handling process which prescribe a spiking of samples in ACE2-HEK293T/A549, VERO E6 and Human lung xenograft mouse.
The “lineages” they claimed to have found aren’t mutational lineages at all—all the mutations they see on these sequences were unique to that specific sequence, and are the result of RNA degradation and from the Taq polymerase errors accumulated from the nested PCR process
Where the extremely low error-ridden cross-contaminating molecule were amplified with mutagenesis happening on each step—especially between the initial RT-PCR and sequencing PCR process, which cross contamination have happened. Notice that any extra Pro or Cys on the highly
Conserved N protein sequences are lethal to the virus—especially the extra Cys which will become a nucleophile and a Ribonuclease in the budded virion in an extracellular environment, degrading the genome and inactivating the virus immediately. These Cys will also cause the N to
Dimerize and consequently lose it’s ability to bind RNA or mediate virion assembly in the first place. While the Pro will break a conserved secondary structure in the C-terminal domain and prevent the N from folding correctly. These are not found anywhere else and are obviously
Sequencing errors. Especially since these samples came from cross-contamination without a corresponding positive oral or anal swab (they didn’t get a single swab that is positive).
M1, they cite as having “hemorrhage” claimed to be Coronavirus symptoms, are anything BUT coronavirus symptoms. https://t.co/3a8nqY6rOz “Open in new tab
X-ray tests revealed a large area of shadow in the left lung of pangolin 2-Lishui, suggestive of pneumonia” but
“we have not found coronaviruses in these pangolins by meta-transcriptomic and PCR methods.”. Notice the PLT- in the Pestivirus-infected pangolin samples. Pestivirus is seen in the SRA of M1, SRX7756769 (107267359 spots as in the Xiao et al article). The Pestivirus is seen
In all PRJNA573298 sample lungs, and it causes interstitial pneumonia and gives the same kind of CT scan and blood test result as in the 06/22 article. This sample also contained a locally circulating CCoV (Canine Coronavirus, often infects zoo/rescued solitary animals due to
It’s ubiquitous nature in most human-associated environments) strain with reads that weren’t rolling circles as the GD-1 reads and are mostly related to the Canine Coronavirus isolates circulating in dogs in GZ.
Coronavirus infections can not generate rolling circles as they does not encode RNA ligases, an enzyme that is necessary to create a true CircRNA. (What they claim to be CircRNA reads from CoV infections in other papers aren’t true CircRNA but are Partially duplicated RNA due to
The Coronavirus RdRp template-switching to a location before it’s current location. This RNA is still linear. However, the Rolling Circles corresponds to two hotspots of M13 primers seen in the amplicons for nsp3 and for the 3’-end of the genome. This imply that they were
Remnants of cDNA cloning, stemming from DNA that were circularized without a vector backbone.)
Canine Coronavirus and FIPV (alphacoronavirus 1) infections also generate very similar symptoms, and will result in positive cross-reactive NP in the immunofluorescence screening. Even the right GEO profile provide they remove it from the final SRA. Finally, the ACE2 expression
“Lowest in the lungs” claimed in this paper is at odd with the 06/22 paper claim that “ACE2 and TMPRSS2 is most highly expressed in the lungs”. This suggest that the GEO profile of the 06/22 article associated with A22/SRX8582289 have been fabricated or were derived from hACE2
Mouse. The hemorrhage seen in M1 is consistent with DYPV(Flavivirus/pestivirus related to BVDV) which is seen in the metagenome, while the pneumonia could be either be the result of DYPV-related Pestivirus, from the Sendai (murine respirovirus), from the Canine Coronavirus or
From any of the non-Coronavirus (Colitivirus or Murine Respirovirus) agents found in PRJNA573298.
M1 have Murine Respirovirus.
https://t.co/ZJnLSxwe5o
SeV is in the same family as human Parainfluenza virus. It causes bilateral ground-glass opacity.
https://t.co/NeY2ajwsDo
The blood tests—pestivirus once again fully explains all the blood changed observed. When a Parainfluenza virus is also present, the blood test become indistinguishable.
Another snRNA-seq say ACE2 and TMPRSS2 are barely exiressed in the lungs and are never coexpressed.
Weaker than the kidneys—this is again at odds with the SCAU claim that even negative pangolins have lung>>kidney for ACE2. Both publication say that Kidney is the highest ACE2 expressing organ in all pangolins where lung expression is negligible. The 06/22 article is the only
One that claim that all pangolins including negative samples, have lung ACE2 >> Kidney. This indicate that they have Spiked all of them with K12-hACE2 mouse as it is in disagreement of all previous or later pangolin transcriptomes.
“(Individual number: L08) as a positive control”—and guess what L08 actually is—Human AND Mice!
L08: Lung08.

More from Daoyu

@JackDempsey2_2 @pathogenetics @ggronvall

@pathogenetics @ggronvall And 100% of all the alleged market animal photos were taken in 2017.
https://t.co/Vh4dKy6rvI there is a reason why that animal paper refused to release the per-month data of animal sales in the wet markets—the sales were completely banned in 2018 when the consumption of

@pathogenetics @ggronvall Contraband animals were prohibited in an 2017 revision fo the CCP’s own “wildlife protection law”. This targets the main reason for consumption, which is to brag as a social status. No animals or their meat were ever sold in wet markets or online after that date. The only things

@pathogenetics @ggronvall Ever sold after the date were desiccated and sun-dried parts for use in jewelry and for medicine. They are always thoroughly dried out which would inactivate all enveloped viruses incl. all coronaviruses within them if they were present.

@pathogenetics @ggronvall https://t.co/2679qQiA0P
The claimed market origin is undermined by the presence of consistent positive SARS-CoV-2 detection in wastewater and in patient samples outside China in at least two different countries well into November 2019, which is before the first case in the wet
@franciscodeasis https://t.co/OuQaBRFPu7
Unfortunately the "This work includes the identification of viral sequences in bat samples, and has resulted in the isolation of three bat SARS-related coronaviruses that are now used as reagents to test therapeutics and vaccines." were BEFORE the


chimeric infectious clone grants were there.https://t.co/DAArwFkz6v is in 2017, Rs4231.
https://t.co/UgXygDjYbW is in 2016, RsSHC014 and RsWIV16.
https://t.co/krO69CsJ94 is in 2013, RsWIV1. notice that this is before the beginning of the project

starting in 2016. Also remember that they told about only 3 isolates/live viruses. RsSHC014 is a live infectious clone that is just as alive as those other "Isolates".

P.D. somehow is able to use funds that he have yet recieved yet, and send results and sequences from late 2019 back in time into 2015,2013 and 2016!

https://t.co/4wC7k1Lh54 Ref 3: Why ALL your pangolin samples were PCR negative? to avoid deep sequencing and accidentally reveal Paguma Larvata and Oryctolagus Cuniculus?
https://t.co/SvP0tfbDki


https://t.co/k9mmL9e5kn


https://t.co/7fR13yb3qJ


https://t.co/EXerlaiG9u


https://t.co/iNCdLY1Pzn
@franciscodeasis https://t.co/Sd9IslUCH5 a FCS need a FCS in the inoculum to exist. It can not arise de-novo as it will be destroyed instantly by the immune system.

https://t.co/UgXygDjYbW a fourth Sars-like CoV is live at the WIV. This fourth virus is an infectious clone, where engineering of the S1-S2 is used regularly as mean to generate a culturable virus in HAE cells. No VERO E6 here, and HeLa-hACE2 is the new VERO of the WIV

https://t.co/DtjyycKy1v
https://t.co/PG7LVnHfsy
Even with VERO E6, only half the time does passage lead to the loss of the FCS—smaller plaques need to be explicitly picked for that to be a certainty.

Marburg virus is a novel virus that escaped from the lab. https://t.co/OGQM6qV27l the only reason why it did not become a pandemic is due to it being too lethal to sustain asymptomatic transmission in humans.

The highest reported number of cases were in WuChang right on top of the old WIV headquarters, In contrast to the population density data of Wuhan—note that the place near the market had the highest population density in all of Wuhan, which make it the most optimal location for

More from Science

@mugecevik is an excellent scientist and a responsible professional. She likely read the paper more carefully than most. She grasped some of its strengths and weaknesses that are not apparent from a cursory glance. Below, I will mention a few points some may have missed.
1/


The paper does NOT evaluate the effect of school closures. Instead it conflates all ‘educational settings' into a single category, which includes universities.
2/

The paper primarily evaluates data from March and April 2020. The article is not particularly clear about this limitation, but the information can be found in the hefty supplementary material.
3/


The authors applied four different regression methods (some fancier than others) to the same data. The outcomes of the different regression models are correlated (enough to reach statistical significance), but they vary a lot. (heat map on the right below).
4/


The effect of individual interventions is extremely difficult to disentangle as the authors stress themselves. There is a very large number of interventions considered and the model was run on 49 countries and 26 US States (and not >200 countries).
5/
Epic thread incoming:

I'm going to answer the question so many people have been asking this week:

WHAT IS PROJECT X???

Here's the definitive thread to tell you - and show you -precisely what Project X is

Grab a drink, sit down with me and let's #TalkLiberation

<3

1/?

"Project X" is actually called "PanQuake".

Pan means "all". Quake is the huge effect our voices can have when our communications are uncensored and when we have access to brand new functionality that *enhances* our social reach, rather than diminishes it

Here's our logo:

2/?


You can follow the fledgling official PanQuake Twitter account here: @pan_quake and see our super cool new website here:
https://t.co/F7wLSeM6aK

You can find our donation page here: https://t.co/VICFnsR0RX

Keep reading this thread to find out why we created it & what it is

3/?

SPOILER ALERT: Much of the content below this point is from my personal slides & speech notes from today's launch event. That stream got totally ruined by (big) tech problems, but I'm happy to report everything is turning out wonderfully


Here are some of our most high profile & dedicated public advocates for PanQuake - many of whom were scheduled to appear at our launch. All of whom stuck around for hours, to do a prerecord of the event, which is being edited, processed & uploaded for you as I write this.

5/?

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