Steve Massey Profile picture
Mar 21, 2023 12 tweets 4 min read Read on X
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

Nov 2
'Conincidentally' we showed pUC57 was also used to clone a genome fragment from an unpublished pangolin-related SARS2r-CoV from China 🧵

h/t @humblesci @Daoyu15 @ydeigin @quay_dr

mdpi.com/2673-8007/2/4/…

x.com/Engineer2The/s…
2/ This striking conincidence indicates that the novel pangolin-related coronavirus we identified from contaminated datasets (GX_ZX45r-CoV) was likely part of an infectious clone (IC), given that one of the reads was ligated to pUC57

From our paper: Image
3/ This can be inferred given that ZLS et al report using pUC57 for cloning genome fragments of HKU5 coronavirus prior to assembly into an IC in their recent Nature paper on engineering coronavirus receptors

h/t @Engineer2The Image
Read 6 tweets
Sep 18
I have published a paper on my analysis of the RaTG13 dataset

RaTG13 is the closest phylogenomic relative of SARS2

Transcriptome and GO enrichment analysis indicates the likely source of the dataset is a bat mating plug, rather than a fecal swab 🧵

mdpi.com/2036-7481/15/3…
A mating plug provenance resolves the long standing conundrum over the low amounts of bacteria in the RaTG13 sample, noted by @MonaRahalkar and others, which is inconsistent with a fecal swab as reported by the WIV, who collected the sample
I presented the evidence for a mating plug provenance in a previous thread 👇
Read 10 tweets
Sep 8
A leak of polio from a research facility is indicated by a poliovirus genome sequence generated from a sample collected in 2014 in China by the Wuhan Institute of Virology

The sequence is 99 % identical to that of a polio strain from 1954 🧵
academic.oup.com/ve/article/10/…
2/ Chesnais et al sequenced 3 poliovirus genomes recovered from 60 year old historic samples generated by Albert Sabin at the Pasteur Institute, Paris

Sabin is the father of the oral polio vaccine, created from attenuated polio strains
Image
Image
3/ Poliovirus is a single stranded positive sense RNA virus, that only naturally infects humans (no other natural hosts are known)

There are 3 poliovirus serotypes: PV-1, PV-2 and PV-3 Image
Read 28 tweets
Jul 26
Why are virologists freely allowed to anonymize lethal synthetic viruses, but developers are put in jail for writing code that anonymize bitcoin transactions ?

Genetically enhanced infectious clones present a much higher risk than anonymous monetary transactions 🧵 Image
@R_H_Ebright @SenGaryPeters @COVIDSelect @BiosafetyNow @CharlesRixey @HSGAC_GOP @RepBradWenstrup @RepRaulRuizMD Virologists often synthesize infectious clones (ICs), which are used to produce live infectious viruses

To make an IC of a coronavirus, due to its large size constituent fragments need to be synthesized, and then ligated together to form a complete genome
Unique restriction (cut) sites at the ends of the fragments allow them to be assembled in the correct order

These are often left in the genome as a signature of the ligation, as in this SARS1-related IC by ZLS and Daszak

journals.asm.org/doi/10.1128/jv…
Image
Read 29 tweets
Jun 25
Unravelling the RaTG13 Dataset's Origin

The RaTG13 dataset has overexpressed transcripts which indicate it was mis-sampled from a bat mating plug

This provides a reasonable explanation for anomalies in the dataset, described as a 'fecal swab' 🧵 Image
2/ RaTG13 is (still) phylogenomically the closest coronavirus backbone to SARS2

There has been a longstanding controversy regarding the origin of the RaTG13 dataset

RaTG13 is described as being generated from a Rhinolophus affinis (Intermediate Horseshoe bat) fecal swab Image
3/ However, the dataset has few bacteria which is inconsistent with a fecal swab, given that fecal material is dominated by bacteria

Read 47 tweets
Jun 21
Re-watching this video, it becomes clear why virologists have lobbied so hard to detach the names of novel pathogens / strains from the location where they arise

"the disease is the same name as the lab" 😆
Detaching regional / place names from pathogens is a form of anonymization eg delta, omega etc

Why would virologists want to use names which are arbitrary and non-informative ? geographic information is an important part of epidemiology and helps understand origin and spread
Claiming that the use of place / regional names is 'racist' is a red flag - geography is neutral

The implication that the uneducated masses who use such terminology are reflexively bigoted is surely a form of prejudice and elitism in itself
Read 4 tweets

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