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Freedom Requires Live-Dead analysis: Subgenomic RNA
You need to pay attention to Wolfel et al and Kim et al.
A very elegant method for Live-Dead analysis.
But before we dive into that Mol-Bio hole, you must see the gravity of Liotti’s work.
https://jamanetwork.com/journals/jamainternalmedicine/fullarticle/10.1001/jamainternmed.2020.7570?guestAccessKey=26a817a5-2e73-4c00-9fc8-42493feb1b6e&utm_source=silverchair&utm_medium=email&utm_campaign=article_alert-jamainternalmedicine&utm_content=olf&utm_term=111220
You need to pay attention to Wolfel et al and Kim et al.
A very elegant method for Live-Dead analysis.
But before we dive into that Mol-Bio hole, you must see the gravity of Liotti’s work.
https://jamanetwork.com/journals/jamainternalmedicine/fullarticle/10.1001/jamainternmed.2020.7570?guestAccessKey=26a817a5-2e73-4c00-9fc8-42493feb1b6e&utm_source=silverchair&utm_medium=email&utm_campaign=article_alert-jamainternalmedicine&utm_content=olf&utm_term=111220
Liotti (&Wolfel) demonstrate that your infectious stage is above 1-2M RNA copies/ml and usually lasts 5 days. However non-infectious RNA can last 77 days. To be safe they suggest 10 days and 100,000 copies/ml as non-infectious.
So what is creating the long tail of qPCR positivity that is not infectious.
This is believed to be due to subGenomic RNA. Kim et al has the best review on this.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7179501/pdf/main.pdf
This is believed to be due to subGenomic RNA. Kim et al has the best review on this.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7179501/pdf/main.pdf
Before we do the deep dive on Subgenomic RNA, Have a look at how Wolfel et al used this to track Infectious vs non-infectious (Live-Dead) SARs-CoV-2.
Wolfel https://pubmed.ncbi.nlm.nih.gov/32235945/
Wolfel https://pubmed.ncbi.nlm.nih.gov/32235945/
The Virus makes 29Kb genomic RNA to be packaged. But it also makes 9 subgenomic RNAs that do not get packaged. These fragments have a leader sequence on their 5' end that can be targeted with an independent qPCR assays so one can measure total gRNA/SubgenomicRNA.
Wolfel exploits this to create a Delta CQ or the difference between live-dead qPCR signals. They then do a very thorough job tracking this with cell culture and Antibody studies and its a wealth of information.
This once again highlights the lack of infectability with late CQ
This once again highlights the lack of infectability with late CQ
So how many people will tolerate an 11+ week quarantine. I hope zero but I'm worried they are boiling frogs and people are getting conditioned to Ivory Tower rulership.
Given there are published methods to estimate infectiousness, what should we do?
Given there are published methods to estimate infectiousness, what should we do?
Live-Dead blind qPCR cannot be the sole data point to classify a case or steal someones liberty. Given the long persistence of qPCR signal seen in Liotti, much of the positivity rise we are seeing today may be seasonality + testing Vol + subgenomic RNA detection.
While cases are on the seasonal rise, excess deaths are not rising as fast as the spring. Flu is gone and likely reclassified as C19 given the endemic nature of its spread in hospitals.
The responsible thing to do for society is gRNA/subgRNA qPCR to understand infectiousness.
The responsible thing to do for society is gRNA/subgRNA qPCR to understand infectiousness.
It's critical we push for this as the economic ruin of lockdown is immense. It is a 1st world privilege that starves the 3rd world to satiate a glutinous fear. A fear so irrational and ungrounded in trade-offs that we need to back up and think globally. https://www.npr.org/sections/coronavirus-live-updates/2020/07/13/890398347/u-n-report-says-pandemic-could-push-132-million-people-into-hunger
You may be tempted to make a blunt CQ cut off with the existing tests. The problem with this is that it's not a very sharp knife. Infectious vs non-infectious patients sit on both sides of any chosen cut off and this defaults into a position of caution. This can sharpen the knife
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