Daoyu Profile picture
10 Feb, 16 tweets, 6 min read
Warning: animals may seroconvert from already-circulating human SARS-CoV or SARS-CoV-2 in the local environment if samples are collected after the widespread circulation of these viruses at the site of collection. Especially captive
Animals as both batches of samples are. All samples are PCR negative and no genome of Coronavirus was obtained from their pangolin samples. Only bats which harbor only useless SL-CoVs that don’t bind ACE2 or have a FCS. nature.com/articles/s4157…
Assembly of the virus samples they provided revealed that the 5’-UTR were all inverted on the final assembly.
This indicate synthetic virus with bat Spike. bat Spike is useless, and they have no inserts. They also invalidates the claim of a PAA insert on RmYN02(NSPVARVG aligns to HTVSLLRSVG as NSP-VAR-VG).
2003 sera was actually collected in Guangdong China, indicate captive exposure to SARS-CoV. Both samples are negative for RT-PCR using the pan-CoV primer (PREDICT primers) indicate that while pangolins can be vaccinated against SARS-like RBD/RBM through exposure to the human
Virus, they does not support infection by these RBM/RBDs. (Samples are all negative for Coronaviruses). This is in agreement with the discovery that pangolin ACE2 bind poorly to these RBDs and that pangolins express ACE2 poorly in all tissues, preventing the establishment of
Productive infection and prevent PCR positivity. The false positive rate from cELISA and sVNT analysis can not be overlooked. ELISA more specifically, suffers from non-specific binding by polymerized proteins in old sera.
Another possibility that can’t be overlooked is the possibility of counterfeit results as there is a relatively low technical barrier against ELISA and sVNT counterfeiting. You just take the civet and the rabbit sera and mix them with the bat and the pangolin sera. Make sure you
Use negative PCR samples or it will just reveal the human/rabbit, or the human virus which indicate infection post-outbreak from human patients.
“PCR testing using pan-CoV primers was conducted on the limited pangolin samples in both studies and were all negative.” Because they ARE FAKE. Using rabbit and civet sera for a spike-in. Those ELISA and sVNT don’t distinguish between real and contamination from other samples.
Digest NA with RNAse and DNAse in rabbit sera and then extract the circulating antibodies using protein A affinity or remove the RNAse and DNAse by reverse affinity purification using the his-tag on the expressed proteins. They have negative PCR for sera so they could pass for
Not sequencing them, which would reveal that the sera have been mixed with rabbit or human immune sera, or that the human virus may have entered the animals during sampling (samples collected after SARS1/SARS2 circulation in humans at that location) and caused the seropositivity
During captivity. They likely functioned as live-attenuated vaccines, as they failed to establish an infection yet is sufficient to cause antibody response.

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More from @Daoyu15

10 Feb
Bat sampling yield useless SL-CoV, pangolin sampling too late to exclude seroconversion through exposure to the human virus, and yielding no viral genomes nor isolates. They just use this as leeway to sneak in fake results. No sequence mean no way to ImageImageImageImage
@michaelzlin Verify the authenticity of either the sera or the alleged “precursor” “similar strain”. Human SARS-CoV and SARS-CoV-2 already begun widespread circulation at specified location BEFORE both sampling takes place. pubs.acs.org/doi/pdf/10.102… environmental persistence lead to exposure of
@michaelzlin Many animals in both case, which lead to seropositivity without infection as the human strain is specific adapted to humans, and fail to establish proper infection in animals. ImageImage
Read 5 tweets
10 Feb
@thekublakhan archive.is/XMloy
Samples taken after the widespread circulation of the human strain are of no forensic value for origin tracing or “evidence of circulation in wildlife”. Animals are captive before sampling and exposure to the human virus is evident in both case. They are
@thekublakhan Vaccinated by exposure to the human virus without infection, as “PCR testing using pan-CoV primers was conducted on the limited pangolin samples in both studies and were all negative.” Indicating that the human virus did not replicate in these animals after exposure, and no
@thekublakhan Reservoirs or circulation exist in these animals (you need to have the live virus to “circulate”, which in this case there is none, as no Coronaviral NA can be detected from them.
Read 5 tweets
9 Feb
@phylogenomics Serological testing are completely useless if you collect them after the local human have already been infected by the pathogen you are testing for. May 2003 is after SARS-CoV have entered Thailand. Mar-Jun 2020 is after SARS-CoV-2 have entered thailand. Wherever you have a lot
@phylogenomics Of human infection, you are bound to have environmental pervalence of the virus and that is going to seroconvert many animals nearby just like a live attenuated vaccine. It does not matter whether the animal is a successful host or not. Even failed infection through poor or
@phylogenomics Limited replication is sufficient to cause Ab positive just like a polio pill or a flu shot.
Read 8 tweets
9 Feb
@wormmaps You are going to have a VERY BAD DAY. All wildlife samples collected after the beginning of SARS-CoV-2 may become contaminated by it from the human source. Animals, even non-susceptible animals, can acquire immunity from a LAV-like effect from the unadapted human virus. Only sera
@wormmaps Collected before the human outbreak are useful for forensic purposes. Coronaviruses have a very broad host tropism and animals that are not otherwise suceptible (they use rabbit sera for cross-Spike neutralization/binding) can still become vaccinated in the same way you get a flu
@wormmaps Shot up the nose.
Read 12 tweets
9 Feb
The P2S dataset is scrubbed clean. All reads that display fusions fuses to bacterial 16S rRNA, bacterial chromosomes and sequences from diverse Eukaryotes and Bacteria. Notably, one read display fusion to a Human BAC. the Homology on other sequences are far lower here.(1-20).
No genus-specific definitive phylogenetic mapping (other than bacterial 16S) could be obtained from any of these reads as the fusion overhangs all have 100% perfect matches to organisms of all kingdoms of life. they really DID a thorough job cleaning it up such that
Read 9 tweets

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