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N-acetyl-L-cysteine to disrupt mucus and increase RNA extraction and sensitivity

A: Not for current challenges

A paper from 2004 showed addition of N-acetyl-L-cysteine to the sample could disrupt the mucus and increase the yield of RNA by ~2.75 times.

Have you validated this method, if so, how and what were the results of the validation?​

Paper here: https://www.ncbi.nlm.nih.gov/pubmed/15324540

"Taking patient ID17 as an example, S = 4,790 copies (958 x 5), M = 8,402 copies (4,680 x 2 – 958 x 1), and C = 330 copies (8,460 x 0.039)."

Where S = saliva supernatant, M = supernatant treated with N-acetyl-L-cysteine, C = cellular fraction.

How quickly could this be deployed and what are the dependencies?​

Very quickly, no dependencies.

What is the likely production volume?​

N-acetyl-L-cysteine is widely available.

What are the risks and barriers to using this at scale?​

Low / none.

Who are you already partnering with on this?

No one yet, might try to encourage OpenCell to perform an experiment.

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Alexander James Phillips Apr 23, 2020

I appreciate this will need to be marked as 'E' (or 'A') as not RNA extraction.

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Bev Matthews Apr 23, 2020

The idea has been progressed to the next milestone.

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Bev Matthews Apr 23, 2020

Status label added: B

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Bev Matthews Apr 25, 2020

Hello Alexander, your solution has been reviewed and the reviewers have asked if you could come back when you have more information and data. Bev

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Bev Matthews Apr 25, 2020

The idea has been progressed to the next milestone.

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heidi Langley Apr 28, 2020

waiting for answer

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Alexander James Phillips Apr 28, 2020

Hi Heidi, Thanks for your message. Who are you waiting for an answer from? What was the question please? Thank you.

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Alexander James Phillips Apr 30, 2020

Hui He et al. 2017 ( https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5362898/ ) show 2M GITC and 80 mM DTT ( Dithiothreitol ) yielded low Ct values for RNA and DNA detection.
A three fold reduction in GITC use would reduce pressure on current supply problems.
Does not assess viral inactivation.
Supernatant is discarded, which given the 2004 paper in this post, is not advisable.

Also there are many other mucolytics that might be used to increase available RNA and assay sensitivity: N-acetylcysteine, erdosteine, fudosteine, gelsolin, thymosin β4, dextran and heparin: https://pubmed.ncbi.nlm.nih.gov/17716385/ https://err.ersjournals.com/content/19/116/127#sec-12

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Sonia Bordin-Aykroyd 10 months ago

Hi Alexander. Just out of interest, can those substance disrupt biofilms? if so, are they bio-compatible? (as, can they be used in the mouth?)

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Alexander James Phillips 6 months ago

Hi Sonia, I am not sure if they can disrupt biofilms but there are certainly members of this class of compounds which are used as pharmaceuticals already so I expect some would have indications for use in the mouth.

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Tom Jordan Jul 23, 2020

Thanks as always for sharing this with us Alexander. The team recognises that there are some labs taking this novel approach for formulating locally to disrupt mucus. In light of where we've now in terms of supplies and what labs are likely to procure or use, we will close this proposal. Though thanks again, we are grateful for this, and the protocol remains here for anyone with an interest in pursuing this.

Tom

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Tom Jordan Jul 23, 2020

Status label added: A: Not for current challenges

Status label removed: B: One to watch

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