Steve Massey Profile picture
Mar 21 12 tweets 4 min read
So, Flo Debarre et al’s raccoon dog analysis that has caused such a media frenzy has been released and what does it show ?

Not much 🧵
2/ Essentially, it confirms Gao et al’s preprint analysis that there was nuc acid from animals in addition to humans in the samples 👇 (no surprise there)

It adds some species specific info
3/ The analysis is crude, and taxonomic attribution method naïve

They rely on seq assembly, which miss a lot of info

They use numbers of assembled contigs as a metric for quantity of species specific material

This is semi-quantitative at best (due to the vagaries of assembly)
4/ Even then, they fail to see if the species specific material correlates with numbers of SARS2 reads, which is inexplicable
5/ They apparently identify a potentially new subspecies of raccoon dog – this would worth following up for forensic purposes, but bafflingly they fail to do so
6/ But they also seem to identify new subspecies of masked palm civet, hoary bamboo rat, Malayan porcupine and Amur hedgehog. This could be interesting, or more likely indicates problems with their assembly. Oddly, they fail to follow this up and validate their assembly qualities
7/ Bizarrely, they seem unable to differentiate between DNA and RNA (hint, trying mapping reads to annotated mito genome or nuc genome). This has importance as it can affect estimates of relative species proportions
8/ Tellingly, none of the stalls with raccoon dog nuc acid have a human SARS2 case linked to it (which puzzlingly they fail to mention)

(figure on left from our HSM Zoonosis critique, on right from Debarre et al)
9/ This saga is a case study in the perils of making grandiose claims without having completed the analysis, and of the hubris to embark into a new subject area without specialists (metagenomic) to scrutinize and suggest robust analyses
10/ The report can be found here:
zenodo.org/record/7754299…
8/ (that disappeared above)

Tellingly, none of the stalls with raccoon dog nuc acid have a human SARS2 case linked to it (which puzzlingly they fail to mention)

(fig on left from our HSM Zoonosis preprint, on right from Debarre et al)

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More from @stevenemassey

Mar 21
9/ This saga is a case study in the perils of making grandiose claims without having completed the analysis, and of the hubris to embark into a new subject area without specialists (metagenomic) to scrutinize and suggest robust analyses
10/ The report can be found here:
zenodo.org/record/7754299…
Image
Read 4 tweets
Feb 22
Here is a short summary of our recent discovery of a novel MERS-related coronavirus clone from Wuhan
We identified an infectious clone of a novel MERS (Middle Eastern Respiratory Syndrome) related coronavirus (MERSr-CoV) in sequence datasets from Wuhan, China. @WashburneAlex provides his perspective here: alexwasburne.substack.com/p/an-unreporte…
Infectious clones are synthetic DNA constructs that are used to ‘revive’ coronaviruses whose genomes have been identified in metagenomic sequencing datasets, usually from bat samples. They do this by allowing the expression of the virus RNA in a host cell
Read 6 tweets
Feb 21
We have uncovered evidence of undocumented GOF work in Wuhan 🧵

We have identified an infectious clone (IC) of a novel MERS-related coronavirus in rice sequencing datasets from Huazhong Agricultural University (HZAU), Wuhan

biorxiv.org/content/10.110…
2/ The novel coronavirus (CoV) is most closely related to merbocovirus HKU4. Merbecoviruses are betacoronaviruses that include MERS-CoV, HKU4-CoV and HKU5-CoV, from bats. We term the novel CoV 'HKU4r-HZAU-2020' to reflect its phylogenetic affinity, place and date of sequencing
3/ HKU4r-HZAU-2020 is contained within a large insert expression vector and represents an IC. The intended cell line is not revealed in the data sets, as the construct is in DNA form and so not transfected and expressed
Read 15 tweets
Nov 30, 2022
Bat Forensics !! 🦇

Horseshoe bats are likely the ultimate reservoir of the ancestor of SARS2. Here I take a closer look at the Rhinolophus bat species present in the Mojiang mine, and discover two potentially new species/subspecies – with implications for the ID of RaTG13
2/ What is the reason for studying these bat species relationships ? Well, RaTG13 and RaTG15 were both isolated from the Mojiang mine, and both were described as being sampled from Rhinolophus affinis, the intermediate horseshoe bat 👇
3/ The low level of bacteria in RaTG13 has been a puzzle for many investigators. Although it is described as being from a R.affinis fecal swab, the low proportion of bacterial sequences present are not consistent with that (fecal material has high numbers of bacteria)
Read 30 tweets
Nov 21, 2022
Michael, I'd be pleased to do a whirlwind tour of the iconic RaTG13 and why the clade 7896 viruses might hold the key to its true identity
RaTG13 is the closest related genome to SARS2 and was published early in the pandemic by the Wuhan Institute of Virology (WIV). A major difference between the two viruses is that SARS2 has a furin cleavage site (FCS), critical for its pandemic potential
The insertion of the FCS in SARS2 compared to RaTG13 led Robert Garry to say that 'I just can't figure out how this gets accomplished in nature'

(strange to tell the first Lab Leak proponents are now fervent zoonosis proponents !)
Read 10 tweets
Nov 21, 2022
Low levels of bacteria in the 'rectal swabs' used to generate 7896 clade sarbecovirus genomes 🧵

On the trail of the true identity of RaTG13 I decided to look at the 7876 clade NGS datasets (which include RaTG15), as these were also generated from the Mojiang mine.... Image
2/ RaTG13 has become iconic in the C19 Origins debate because phylogenomically it is the closest relative of SARS2, but there are anomalies and opacity in its history and sequencing
researchgate.net/publication/35…
3/ Of great interest, it came from the Mojiang mine in Southern China, where 3 miners died of a potential SARS-related illness h/t @MonaRahalkar @BahulikarRahul @TheSeeker268
frontiersin.org/articles/10.33…
Read 18 tweets

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