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
Pekar et al have published a paper claiming that 2 SARS2 A/B intermediates from Feb 2020 are derived rather than ancestral
The existence of ancestral intermediates would falsify Pekar et al's earlier claim of 2 independent spillovers of lineages A and B at the Huanan Market 🧵
2/ Hence Pekar et al undergo extraordinary convolutions in order to try and claim the 2 A/B intermediates, identified by Lv et al 2024, are late arrivals rather than the original intermediates that led to the evolution of lineage B from lineage A
3/ To sum, they once again (ab)use modeling to support intuitively unlikely scenarios
They claim it is more likely that the 2 Lv et al intermediates arose from either Hu-1 (lin B), or its closest lin A relative (C8782T,T28144C) AFTER the 2 lineages had originally split
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:
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
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 👇
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
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 🧵
@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