Search for a Biomarker: Ron Davis interviewed on ME/CFS Alert

Ben H

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Hi guys,


Dr. Ron Davis Talks with Llewellyn King about ME/CFS Research

Llewellyn King, host of ME/CFS Alert, has just released his newest episode interviewing OMF Scientific Advisory Board Director Ronald W. Davis, PhD. This edition focuses on the critical research led by Dr. Davis to identify a biomarker for ME/CFS. This is the first in a series of interviews that were recorded at the OMF sponsored 3rd Annual Working Group meetings and Community Symposium at Stanford University. Watch Llewellyn's interview with Dr. Ron Davis. And stay tuned for additional episodes featuring OMF's team coming out soon.

Watch ME/CFS Alert: Finding an ME/CFS Biomarker, Ronald Davis, Stanford University - ME/CFS Alert #109 here.






Support research to end ME/CFS.


www.omf.ngo



B
 
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Interesting. Dr. Davis still sounds pretty optimistic about the Metabolic Trap. In addition, it sounds like there is a potential explanation for the 1/70 severe patient in their IDO2 gene study who does not have any damaging mutations. I don’t think that information was shared at the symposium.
 
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Interesting. Dr. Davis still sounds pretty optimistic about the Metabolic Trap. In addition, it sounds like there is a potential explanation for the 1/70 severe patient in their IDO2 gene study who does not have any damaging mutations. I don’t think that information was shared at the symposium.
I agree, I came away from the symposium footage thinking that the IDO trap was further less likely than and this interview has somewhat changed that perspective, especially considering that it is right before the symposium.

I found really interesting the way Ron discussed that normally people recover from disease and that such a bi-stability "trap" if I'm correct in saying could potentially relate to numerous other diseases due to a drastically altered homeostasis. Also, this could be what Dr Phair is alluding to when he is saying the most important work he's done is the trap hypothesis altogether, not specifically the IDO trap. Perhaps he's optimistic that it could have numerous other chronic disease applications - in no way am I putting words in his mouth though.

Ron's discussion of multiple symptom expression due to inherent genetic differences (i.e. we're all different people genetically) is also extremely interesting. It seems logical then that a unifying theory could still be explanatory for disease cause as the difference in symptom expressions among patients - if, what is wrong is an altered homeostasis due to such a trap - could be just due to inherent genetic differences. Also, this perhaps explains the large range of severity too.

Hope that is coherent!
 

gbells

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I honestly have no idea how Dr. Davis' test works. It uses plasma which doesn't contain any cells. So what is it detecting? Have any independent researchers confirmed his findings? It smells very commercial.
 
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I honestly have no idea how Dr. Davis' test works. It uses plasma which doesn't contain any cells. So what is it detecting? Have any independent researchers confirmed his findings? It smells very commercial.
He specified that they measure white cells in plasma, so it is I'm assuming the impedence across the cell, while suspended in plasma. He states that healthy white cells in patient plasma cause similar/same results. Perhaps whatever is in the plasma is interfering with the proceses inside the white cells, thus the measurement is still on the white cells, but now affected by the patient plasma.

Surely he would've done purely plasma tests to rule out that as the signal generator.
 

gbells

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If its using white cells then that makes more sense. White cells would include macrophages and t cells which would be gobbling up viruses and have nuclei that could be infected. Leakage from the mitochondria would increase the positive charge and change the impedance in a measurable way. Large numbers of chronically infected cells vs a normal healthy acute response could be detectable. Clever.

I just read some articles about it. One pointed out that it still has to be validated against different diseases to show it is specific. For example, a patient might test positive if they have a chronic bartonella infection.

“I think the next thing they need to do is test a lot more patients and compare a lot of other diseases — like MS — to make sure what they’re seeing is truly because of ME/CFS,” said Brian Vastag, a former journalist for the Washington Post who has been outspoken about his experience living with ME/CFS.
https://www.statnews.com/2019/04/30/experimental-test-may-help-confirm-chronic-fatigue-syndrome/
 
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nandixon

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@Ben H, Speaking of a biomarker, I may have missed it but I was surprised that nothing was said about the "something in the blood" finding at the recent Stanford Symposium.(?) In particular I was hoping they would either confirm or refute the possibility of an exosome, which @Cort had mentioned back in May:
https://www.healthrising.org/blog/2...e-nanoneedle-scores-big-in-first-me-cfs-test/

If it's true that an exosome still remains a possibility but the team is having trouble isolating the exosomes for testing, would either you or @Janet Dafoe (Rose49) please let Ron know about the following:

This is one of the simplest procedures for isolating exosomes I've seen (using equipment on hand). It can be done in less than 3 hours and only requires an ultracentrifuge (which Stanford should certainly have) and a single filtration step:

Plasma exosomes were isolated by centrifuging whole blood at 600g for 10 minutes, and the supernatant was further centrifuged at 20,000g for 20 minutes. This supernatant was filtered through a 0.22-mm filter and ultracentrifuged at 140,000g for 90 minutes, after which, the pellet was washed in PBS [phosphate-buffered saline].
(The above provides all the necessary info but see the attached PDF for the study this was taken from. [Edit: Actually, it's missing the temperature for carrying the operations out at. I think 4 degrees C is typically used.)

The exosome-containing pellet (from an ME/CFS patient) would then be added to healthy control plasma with healthy control cells and tested in the nanoneedle. We might know very quickly then if an exosome was in fact a likely candidate to be the "something in the blood."

Obviously, an exosome with a specific signature (a surface marker or cargo) could not only make a great biomarker but could also lead to identification of the underlying disease process.

Edit: The obvious caveats are the inclusion of non-exosomal material in the pellet, as well as possible damage to exosomal components during ultracentrifuging. I just thought it would be something potentially very quick that could be done.
 

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FMMM1

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Hi guys,


Dr. Ron Davis Talks with Llewellyn King about ME/CFS Research

Llewellyn King, host of ME/CFS Alert, has just released his newest episode interviewing OMF Scientific Advisory Board Director Ronald W. Davis, PhD. This edition focuses on the critical research led by Dr. Davis to identify a biomarker for ME/CFS. This is the first in a series of interviews that were recorded at the OMF sponsored 3rd Annual Working Group meetings and Community Symposium at Stanford University. Watch Llewellyn's interview with Dr. Ron Davis. And stay tuned for additional episodes featuring OMF's team coming out soon.

Watch ME/CFS Alert: Finding an ME/CFS Biomarker, Ronald Davis, Stanford University - ME/CFS Alert #109 here.






Support research to end ME/CFS.


www.omf.ngo



B

@Janet Dafoe (Rose49) Hi just an update on a European petition for funding for biomedical research for ME. The petition will be discussed at the European Parliament’s Committee on Petitions (PETI) on October 3rd in Brussels. It’s scheduled for discussion around 11.05 CEST [petition no 0204/2019].

You can follow the committee proceedings live by webstream at the following link http://www.europarl.europa.eu/commi...fdDPOay-Xzyvo_xeqqOlyyJNd4vYwYHFhu2BkECspFb3k

If you are unable to watch the meeting live, a recording will be available the following day.

ME Action are supporting the petition; there’s a link to their announcement below*.

Also, readers/friends of readers who are citizens of the European Union (EU) can sign the petition.

You might be surprised how far behind the EU is compared to the US. Lyme disease research received 33.9 million euros (roughly US dollars) from the EU Commission in the last 10 years; ME received nothing.

Research to develop a diagnostic test (e.g. nano-needle) and treatments requires funding; for any readers in the EU please contact your elected representative (MEP) and ask for their support.

*
https://m.facebook.com/story.php?st...29643,"role":1,"share_id":0}]}}&__tn__=*s*s-R
 

FMMM1

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I honestly have no idea how Dr. Davis' test works. It uses plasma which doesn't contain any cells. So what is it detecting? Have any independent researchers confirmed his findings? It smells very commercial.
Haven't read other responses - so you may already have a reply. Basically Fluge and Mella (2016) found that if you added plasma, from a person with ME, to healthy muscle cells then the energy production changed -- the cells seemed to be struggling.
What Ron has found is that if you add salt to the cells then the impedance (electrical resistance) changes dramatically. This seems to be a consequence of a change in cellular energy production i.e. ME cells struggle to remove the salt from the cell -- they can't do the work required.

In terms of other researchers check out Bhupesh Prusty's talk at NIH Conference (April 2019 -- "Accelerating Research--"). Bhupesh found that if you add plasma, from a person with ME, to healthy cells then mitochondrial fragmentation increases. Also, Karl Morten has presented similar data; he measured cellular oxygen consumption rather than electrical resistance/impedance (Ron) or mitochondrial fragmentation (Bhupresh).

It would be interesting to know how Ron's nano-needle data compares with other data e.g. cellular oxygen consumption (Karl) or mitochondrial fragmentation (Bhupresh) or mass spectrometry data.

Also, in a general population classified as having ME how many test positive using the nano-needle?

Short answer is Ron ( nano-needle) may have found a reliable way to find out if people with ME have altered cellular energy production.

I'm not sure you should be worried by commercial e.g. if the nano-needle works for other diseases (MS, Lyme --) then it's easier to develop the technology. Possibly it would be worrying if it's non-commercial; if it is then how can you deliver it (donations/Governments)?

Sorry to add to the information but check out Maureen Hanson's presentation. Didn't watch it all but Maureen's working on exosomes in plasma. These exosomes seem to be responsible, or rather what they contain, for the change in cellular energy production. The exosomes seem to have a signalling compound which changes the behaviour of healthy cells.
 

JES

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Sorry to add to the information but check out Maureen Hanson's presentation. Didn't watch it all but Maureen's working on exosomes in plasma. These exosomes seem to be responsible, or rather what they contain, for the change in cellular energy production. The exosomes seem to have a signalling compound which changes the behaviour of healthy cells.
Yeah, Hanson's presentation I reckon pretty much covers the status on the "something in the blood" finding. To paraphrase it shortly, they have looked into extracellular vesicles (EVs), of which exosomes is one group. They found no change in average size or total concentration of EVs between healthy and ME/CFS groups and neither in individual cytokine levels in EVs. They found, however, that correlations between different cytokines were different in ME/CFS group and healthy controls both in EVs and plasma.

To me it seems this energy metabolism and "something in the blood" research is leading the researchers back to the immune system and abnormal cytokines. If this is the case, I'm not very optimistic it will lead to any major discoveries anytime soon, as cytokines and immune system in ME/CFS has been studied for decades. If it really comes down to the metabolic issues being caused by cytokine signaling, then this isn't the root cause of the disease.

The way I see it, the only finding of OMF that so far comes close at a root cause would be the metabolic trap model. If the trap can be demonstrated to exist in vivo, then great, at least there is a cause found. If not, I'm not sure where they'd go from there. Hanson went on to talk about ME/CFS outbreaks and how a hit-and-run virus could possibly trigger the disease. IMO it would be an absolute key to understand at least if a subgroup of patients have active infections. The research regarding viruses and ME/CFS seems to move along slowly, like in the past 20 years, but I understand it's probably financial limitations again at play for the most part.
 
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FMMM1

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Yeah, Hanson's presentation I reckon pretty much covers the status on the "something in the blood" finding. To paraphrase it shortly, they have looked into extracellular vesicles (EVs), of which exosomes is one group. They found no change in average size or total concentration of EVs between healthy and ME/CFS groups and neither in individual cytokine levels in EVs. They found, however, that correlations between different cytokines were different in ME/CFS group and healthy controls both in EVs and plasma.

To me it seems this energy metabolism and "something in the blood" research is leading the researchers back to the immune system and abnormal cytokines. If this is the case, I'm not very optimistic it will lead to any major discoveries anytime soon, as cytokines and immune system in ME/CFS has been studied for decades. If it really comes down to the metabolic issues being caused by cytokine signaling, then this isn't the root cause of the disease.

The way I see it, the only finding of OMF that so far comes close at a root cause would be the metabolic trap model. If the trap can be demonstrated to exist in vivo, then great, at least there is a cause found. If not, I'm not sure where they'd go from there. Hanson went on to talk about ME/CFS outbreaks and how a hit-and-run virus could possibly trigger the disease. IMO it would be an absolute key to understand at least if a subgroup of patients have active infections. The research regarding viruses and ME/CFS seems to move along slowly, like in the past 20 years, but I understand it's probably financial limitations again at play for the most part.
I've only scanned your post. I think Maureen Hanson found an increase in the numbers of the smallest size - not sure if that's the smallest of exosomes or extracellular vesicles (exosomes?). I intend to try to watch the videos when they are posted online.

It's far from clear what is causing the switch in cellular energy production e.g. cytokines, microRNA's ---. The link to exosomes is simply that they are the correct size i.e. filter out that size category then the effect disappears. Possibly, if Maureen has found a size category which is increased then that might help to identify the signalling compound/parent cells (the cells which produce the exosomes).

I understand that those who know a lot more than I do are cautious about the something in the blood - it needs to be tested by others.

My view is that exosomes --- something in the blood -- are downstream consequence. Robert Phair describes himself as a top down scientist rather than a bottom up --. It would be good to find what is causing something in the blood; but Robert's approach may get there more quickly.

The nano-needle may be a way of identifying drugs/treatments which improve the quality of people lives i.e. as well as diagnosis -- seems reasonable to me. It might get some people to treatments relatively quickly.

I've seen stuff recently on a bug hiding in the system causing Alzheimer's (gingivilas - gum disease). So yes bugs, and virus's, are a possibility in my view. The interest in other diseases (Alzheimer's etc.) may help to develop a model for this latent bug/virus (causing immune activation) theory. I think Bhupresh Pust's work also relies on some post infection effect. Bear in mind Ron Davis's view (talk after Bhupresh's at NIH Conference April 2019) that there's no RNA/DNA i.e. genetic fingerprint to support latent bug/virus theory.

Re funding:
Dr Vicky Whittemore (NIH) at Invest in ME Conference (2019):
"advocacy groups --- that's what makes the difference -
- when they [elected representatives] hear that, from people with the disease -
- advocates -- telling them [elected representatives] what's needed is really what makes the difference"
 

FMMM1

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It can't be from exosomes because they are only pieces of cell contents that can't adapt to stress.
https://bmcbiol.biomedcentral.com/articles/10.1186/s12915-016-0268-z

The previous comment about them containing T-cells makes more sense.
Hanson mention's that exosomes, or rather a microRNA they contain, are a diagnostic for breast cancer. MicroRNAs are involved in the regulation of gene expression - don't ask me how (I'd probably get it wrong). Also, my understanding is that exosomes may contain cytokines.

To me exosomes are interesting; NIH are interested since they are a signalling system - they've helped to understand complex diseases (sound familiar?). How can you be so ill and your blood results look OK (Ron Davis on Whitney) -- exosome signalling?
 
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nandixon

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Yeah, Hanson's presentation I reckon pretty much covers the status on the "something in the blood" finding. To paraphrase it shortly, they have looked into extracellular vesicles (EVs), of which exosomes is one group. They found no change in average size or total concentration of EVs between healthy and ME/CFS groups and neither in individual cytokine levels in EVs. They found, however, that correlations between different cytokines were different in ME/CFS group and healthy controls both in EVs and plasma.
@JES, Hanson's work on exosomes is completely independent of and separate from Ron Davis's finding that upon filtering ME/CFS blood using various size filters that they are (apparently) left with something that has a size consistent with an exosome as potentially being the "something in the blood" that causes the abnormal nanoneedle results when healthy control cells are placed into ME/CFS plasma.

If an exosome can be verified by Ron's group then the idea would be to further use the nanoneedle to attempt to additionally narrow down what exactly the exosome contains or is on its surface that causes the abnormal nanoneedle result. (This could involve not only fractionating the components of the exosome but also fractionating one type of exosome from another type.) The use of the nanoneedle as an analytical tool in this way seems critical to me.

This is in contrast to what Hanson is doing of simply comparing ME/CFS exosomes to healthy exosomes which I'm afraid is not likely to be that fruitful based on decades of prior ME/CFS research.
 
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nandixon

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If an exosome can be verified by Ron's group then the idea would be to further use the nanoneedle to attempt to additionally narrow down what exactly the exosome contains or is on its surface that causes the abnormal nanoneedle result.
Just to clarify this part in case it's not obvious, the exosome itself wouldn't directly cause the abnormal nanoneedle results. Rather it's the effect that the exosome has upon the cell that would (hypothetically) affect the nanoneedle.

As an example, an exosome might have a specific surface component/marker that instructs a cell it comes in contact with to detrimentally change the cell's mitochondrial functioning (permeabilize the inner mitochondrial membrane and create a proton leak, for example). Or it might deliver a peptide to the cell that would detrimentally bind to a mitochondrial enzyme such as ATP synthase (Complex V) and inhibit its function.

The surface marker or peptide would then become the biomarker for diagnosing ME/CFS.
 

FMMM1

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@JES, Hanson's work on exosomes is completely independent of and separate from Ron Davis's finding that upon filtering ME/CFS blood using various size filters that they are (apparently) left with something that has a size consistent with an exosome as potentially being the "something in the blood" that causes the abnormal nanoneedle results when healthy control cells are placed into ME/CFS plasma.

If an exosome can be verified by Ron's group then the idea would be to further use the nanoneedle to attempt to additionally narrow down what exactly the exosome contains or is on its surface that causes the abnormal nanoneedle result. (This could involve not only fractionating the components of the exosome but also fractionating one type of exosome from another type.) The use of the nanoneedle as an analytical tool in this way seems critical to me.

This is in contrast to what Hanson is doing of simply comparing ME/CFS exosomes to healthy exosomes which I'm afraid is not likely to be that fruitful based on decades of prior ME/CFS research.
I haven't watched the presentations; apart from a little of Maureen Hanson's.

I assume that Hanson's NIH funding is, among other things, to look at microRNAs in exosomes. She mentioned that so far they consider that there is no single microRNA which is diagnostic. In terms of causing the change in cellular energy production, I assume that microRNAs are still possible candidates. Is Hanson also studying cytokines in exosomes? I'm not aware of a previous study regarding exosomes in ME; I think Hanson's exosome work, and work by others such as Bhupesh Prusty, has the potentially to help explain ME (or at least a sub-group).

If you can narrow down the size band of the particle which changes cellular energy production (using the nano-needle) then I guess you could do mass spectrometry to identify the proteins and thereby try to identify the parent cells (if it's an exosome) and/or the signalling factor.

Re funding:
Dr Vicky Whittemore (NIH) at Invest in ME Conference (2019):
"advocacy groups --- that's what makes the difference -
- when they [elected representatives] hear that, from people with the disease -
- advocates -- telling them [elected representatives] what's needed is really what makes the difference"
 

nandixon

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In terms of causing the change in cellular energy production, I assume that microRNAs are still possible candidates.
They theoretically should be, but a microRNA's speed of action is, on average, going to be slower than the two possibilities I mentioned (a surface signaling component or an inhibitory peptide) because generally microRNAs affect the formation of proteins/enzymes from mRNA. And I thought that ME/CFS plasma causes healthy cells to give the abnormal nanoneedle signal pretty quickly.(?) If on the other hand they allow the cells to spend any significant amount of time in the plasma before testing then all bets are off.

I'm not aware of a previous study regarding exosomes in ME;
There's definitely at least one study. It may have come from Spain.

Edit: Here it is:
Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5912186/
 
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FMMM1

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They theoretically should be, but a microRNA's speed of action is, on average, going to be slower than the two possibilities I mentioned (a surface signaling component or an inhibitory peptide) because generally microRNAs affect the formation of proteins/enzymes from mRNA. And I thought that ME/CFS plasma causes healthy cells to give the abnormal nanoneedle signal pretty quickly.(?) If on the other hand they allow the cells to spend any significant amount of time in the plasma before testing then all bets are off.


There's definitely at least one study. It may have come from Spain.

Edit: Here it is:
Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5912186/
Thanks.

Based on a few seconds the Spanish study shows more, and smaller, extracellular vesicles in ME. Also the protein profile is different i.e. ME versus healthy controls. Need to listen to/watch Hanson's presentation.

If I recall correctly the nano-needle shows a sudden change. Also, from memory, Bhupesh Prusty found an immediate effect.

Bob Naviaux's description of a cell danger response comes to mind -- what would cause that?

Possibly some clue to the parent cell could be obtained from protein analysis (using mass spectrometry) i.e. if it's an extracellular vesicle.

Could the nano-needle be used to diagnose ME and identify candidate drugs --- if so is that enough i.e. to diagnose and treat some people?
 

nandixon

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Could the nano-needle be used to diagnose ME and identify candidate drugs --- if so is that enough i.e. to diagnose and treat some people?
I think it's unlikely that the nanoneedle could be used to specifically diagnose ME/CFS. I think it may essentially be measuring mitochondrial membrane potential, although perhaps indirectly by measuring the cellular plasma membrane potential. If that's the case, then the nanoneedle is likely to see similar impedance abnormalities in many other diseases, including active MS, SLE, Alzheimer's, Parkinson's, etc., and even autism. (Although the level of impedance might possibly be significantly greater in ME/CFS.)

The nanoneedle could obviously be used to attempt to identify candidate drugs, as they are already doing (e.g., the SS-31/elamipretide finding), but with the very large caveat that a drug that normalizes the nanoneedle may not necessarily have a beneficial effect on ME/CFS symptoms - as I noted previously in a post here:
https://forums.phoenixrising.me/thr...ytes-from-me-cfs-patients.77577/#post-2229686

I think the more important role for the nanoneedle is as an analytical tool to use for narrowing down and finding what exactly the "something in the blood" is.
 

FMMM1

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I think it's unlikely that the nanoneedle could be used to specifically diagnose ME/CFS. I think it may essentially be measuring mitochondrial membrane potential, although perhaps indirectly by measuring the cellular plasma membrane potential. If that's the case, then the nanoneedle is likely to see similar impedance abnormalities in many other diseases, including active MS, SLE, Alzheimer's, Parkinson's, etc., and even autism. (Although the level of impedance might possibly be significantly greater in ME/CFS.)

The nanoneedle could obviously be used to attempt to identify candidate drugs, as they are already doing (e.g., the SS-31/elamipretide finding), but with the very large caveat that a drug that normalizes the nanoneedle may not necessarily have a beneficial effect on ME/CFS symptoms - as I noted previously in a post here:
https://forums.phoenixrising.me/thr...ytes-from-me-cfs-patients.77577/#post-2229686

I think the more important role for the nanoneedle is as an analytical tool to use for narrowing down and finding what exactly the "something in the blood" is.
Interesting.

Lets say that if you have MS and you test positive using the nano-needle; are the other tests (for MS) sufficient to say "you have MS not ME"? I think that if the other tests (MS etc.) are sufficiently accurate then testing positive on the nano-needle isn't a problem. I'm in favour of simply doing a large study of people with ME, and also other diseases which have significant fatigue as a symptom (MS etc.), i.e. using the nano-needle.

Possibly Ron Davis was correct i.e. when he said that a test which showed you are not healthy may have value - i.e. the nano-needle.