The Enterovirus Theory of Disease Etiology in ME/CFS: A Critical Review (O'Neal and Hanson, 2021)

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It's a reasonable statement, but the functional word is "can", not "always". Chia knows how to do it, he's done it in his studies, it's just not something he's equipped to do at scale on a clinical basis. Others have as well (I linked two previous studies that found enterovirus RNA in spinal fluid, not important to read if you are too sick to, just be aware they exist). I can't give you a figure on how quickly, but for example in Chia's previous studies he mentioned he would literally have to take the blood immediately after having it drawn and drive it back to the lab to spin down and process, and even doing so would net very very low copy numbers with qPCR. No commercial or nationalized health system that I'm aware of will use RNA preservatives in the tubes when they draw your blood/CSF for PCR testing, nor will they be guaranteed to process it with any rapidity, nor are you guaranteed that there happened to be any meaningful quantity of cell-free RNA in the few ml of blood they drew.

If enough interest and evidence can build around this idea, I really hope that there will be improvements in clinical testing of enteroviruses so it can be dragged out of the 20th century ideology behind the testing that's barely available even today.
Anyway, my only chance is to give it a try. And I also hope there will be good research coming up on this topic!
 

Pyrrhus

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Is it TRUE or FALSE that enterovirus can be detected in the blood of patients with a persistent enteroviral infection?

I don't think anyone disputes the fact that enterovirus is not detected in the blood using standard laboratory techniques.

And most knowledgable people in the field would not dispute the fact that persistent enteroviruses prefer to spread directly from cell-to-cell, avoiding interstitial fluid, blood, and cerebrospinal fluid.

But:

The other problem is that for a long time people have probably been completely throwing out the [white blood cells] fraction (or at least not testing it) that actually does contain virus in it, the buffy coat. Take a look at the study I linked previously. The virus (or genome fragments thereof) can end up in [white blood cells] which are present in the blood. Co-culturing those [white blood cells] with permissive cell lines can produce a nice, natural amplification effect and can ease [enteroviral] RNA detection. It's not something that will be used anytime soon clinically I imagine, but it's something ME researchers should be paying attention to for sure.
1) @halcyon is referring to this study:
Revealing enterovirus infection in chronic human disorders: An integrated diagnostic approach (Genoni et al., 2017)
https://forums.phoenixrising.me/thr...ders-an-integrated-diagnostic-approach.56060/

Antonio Toniolo, the author of the study mentioned, has developed a technique to co-culture white blood cells with permissive cell lines to make it easier to detect very small amounts of enterovirus in the blood. He has used this technique to identify enterovirus in the blood of patients with post-polio syndrome.

I don't know of any published reports that have tried to replicate Toniolo's technique. Toniolo is now retired and has recently remarked that his students can not get funding to continue this line of research.

But:
From April 2019
Maureen Hanson, is also looking for enteroviruses in the blood samples she gets from the cPET tests. She is trying to culture the samples for over 1 month to see the virus can be grown, similar to what Toniolo did for type 1 DM.

for example in Chia's previous studies he mentioned he would literally have to take the blood immediately after having it drawn and drive it back to the lab to spin down and process, and even doing so would net very very low copy numbers with qPCR. No commercial or nationalized health system that I'm aware of will use RNA preservatives in the tubes when they draw your blood/CSF for PCR testing, nor will they be guaranteed to process it with any rapidity, nor are you guaranteed that there happened to be any meaningful quantity of cell-free RNA in the few ml of blood they drew.
2) Using less advanced techniques than that used by Toniolo, Chia has also identified enterovirus in the blood, simply by using blood collection tubes that contain RNA preservatives and by speeding up processing of the blood. However, I am not familiar with the exact specifics of Chia's technique.

3) Lastly, we must consider the 2019 publication by Charles Chiu in collaboration with the Workwell foundation, which looked at whole blood, including white blood cells, in 14 female patients diagnosed with the Canadian Consensus Criteria:

Whole blood human transcriptome and virome analysis of ME/CFS patients experiencing post-exertional malaise following cardiopulmonary exercise testing (Bouquet et al., 2019)
https://forums.phoenixrising.me/thr...-pem-following-cpet-bouquet-et-al-2019.84824/

Excerpt:
Bouquet et al 2019 said:
Previously, we did not detect evidence of immune dysregulation or virus reactivation outside of PEM periods. Here we sought to determine whether cardiopulmonary exercise stress testing of ME/CFS patients could trigger such changes.

ME/CFS patients (n = 14) and matched sedentary controls (n = 11) were subjected to cardiopulmonary exercise on 2 consecutive days and followed up to 7 days post-exercise, and longitudinal whole blood samples analyzed by RNA-seq.
[...]
We detected sequences from a small number of viruses in the RNA-seq data, including enterovirus A, influenza A virus, anelloviruses/torque teno viruses (TTVs), and human herpesviruses (HHVs) (Table 5).
[...]
The overall virome composition in ME/CFS did not differ significantly from controls (P = 0.746 by chi-square test)
[...]
Reasons for the absence of differential gene expression between ME/CFS patients and controls include
[...]
(3) localization of ME/CFS pathogenicity to a specific tissue (e.g. skeletal muscle or brain tissue) rather than blood
[...]
here, we observed no differences in viral abundance in ME/CFS patients following exercise.
[...]
A limitation of the current study is the small size of the study cohort.
(In this 2019 study by Charles Chiu, the only enterovirus detected in the blood belonged to a control patient, not to an ME patient.)
 
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sometexan84

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Not all of the serotype tests max out at 1:640. Coxsackie B4 for some reason produces a larger humoral response than other enteroviruses and can produce effective antibody concentrations at the 1:1024 dilution.

Don't make the mistake of treating this microneutralization assay as being quantitative in any way. It's a measure of viral neutralization effect in vitro, which then provides an indirect measure of neutralizing antibody concentrations in the blood sample. It can't really inform how much actual viral load is present in the body at any one time.

The presence or absence of enterovirus RNA in those samples at various time periods from the study are much more direct and interesting to look at.
How dare you question me!

jk :D

Yea, I mean, I'm sure you're right, and I'd wager could be even higher than 1:1280 in some cases.

And I could be wrong, but I think ARUP just doesn't test for higher than 1:640, so if they reach that, they put >=1:640. I think that's the case? And Chia uses ARUP, so I thought that max was relevant for those graphs.

But I have no clue as to the max titers from other labs.
 
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That's all very nice. Studies with small groups, theoretical possibilities to detect enteroviral RNA that are completely unrealistic in a clinical setting (at least for people with very severe ME who don't have a Chia but only their GP) and so on...

Furthermore, the Enterovirus Foundation says

“Titers of 1:160-320 and higher are good indicators of current infection.”

@Pyrrhus I don't think that the etiology of ME has anything to do with an ongoing viral infection. But I see that many improve on antivirals. And as long as there are no real treatments one has to make a choice. I'm sure in Germany almost nobody will do a biopsy based on the lab results and the possibility to get a sample for a PCR bc there is no indication to do a stomach biopsy. And the study you cite is really small and has no meaning other than: “not every ME patient we tested has a viral infection.”

So me and most patients are left with the “good indicator” and have to make a choice if they want to give high titers a meaning or not.
 

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I hope I'm not taking this thread too far off-topic, but I wanted to share some gems from the paper mentioned in the title of this thread... ;)

The Enterovirus Theory of Disease Etiology in ME/CFS: A Critical Review (O'Neal and Hanson, 2021)

Introduction
[...]
ME/CFS case documentation shows evidence of both sporadic events involving singular individuals and regional outbreaks involving significant fractions of affected communities, especially hospitals, schools, and military bases.
[...]
The pattern of transmissibility, and acute symptom constellation reminiscent of a flu-like illness, led early investigators to hypothesize a viral theory of ME/CFS disease etiology.
[...]
Between the 1930s and 1960s, a number of globally occurring ME/CFS outbreaks, with a spatiotemporal incidence coinciding with poliovirus epidemics, appeared under the titles of “abortive or atypical poliomyelitis” transitioning to “benign myalgic encephalomyelitis” or “epidemic neuromyasthenia” as physicians sought a term to describe the symptom profile of affected individuals.
[..]
Overall, most epidemic outbreaks have occurred in mid-spring through early fall indicating a virus with seasonal epidemic trends may be involved. Seasonality is not rare for viruses; many types, including but not limited to echovirus, coxsackievirus and poliovirus-related species, are well-known to have strong outbreak seasonality peaking in the month of August or early fall.
[...]
The occurrence of considerable symptom constellation overlaps between ME/CFS, poliomyelitis and other non-polio enterovirus-related clinical outcomes as well as similarity in epidemic seasonality is further circumstantial evidence for a relationship between ME/CFS and enteroviruses.
[...]
To date, the body of research investigating enterovirus [EV] infections in relation to ME/CFS supports an increased prevalence of chronic or persistent infections in several ME/CFS patient cohorts.
[...]
Although a significant number of early papers provided evidence for an association of chronic enteroviral infections with ME/CFS, research into the enteroviral theory of disease etiology largely died out in the early 2000s with a few exceptions. One reason that enteroviral research in ME/CFS has languished is the difficulty of detecting virus after time has passed following an acute infection.
[...]
ME/CFS patients have a number of pathophysiological traits that point to abnormalities in the [Autonomic Nervous System (ANS)], including impaired blood pressure variability, orthostatic intolerance, high prevalence and severity of postural orthostatic tachycardia syndrome (POTS), delayed gastric emptying, impaired thermoregulation in adolescent patients, loss of capacity to recover from acidosis on repeat exercise, abnormal cardiac output and altered brain characteristics in a wide variety of brain regions including the limbic system structures that govern the ANS. [...] Many of the altered brain characteristics seen in ME/CFS patients are similarly reported in clinical cases associated with neurotropic enteroviruses [EVs].
[...]
EVs gain access to the [Central Nervous System (CNS)] through a diverse set of entry mechanisms including direct infection of brain microvascular endothelial cells, retrograde axonal transport following muscle infection, exosomal transport across the blood-brain barrier (BBB), and hitchhiking inside of migratory infected immune cells with BBB privilege. [...] Several known EV CNS infections display autonomic dysfunction symptoms reminiscent of those described in ME/CFS patients.
[...]
Three ME/CFS post-mortem brain autopsy studies found enteroviral genomic RNA and VP1 capsid protein in the hypothalamus, brainstem, cerebral cortex, medial temporal lobe, lateral frontal cortex, occipital lobe, and cerebellum. These findings provide additional support that a persistent EV infection within patient limbic and extra-limbic tissues is possible and could be driving the ANS dysfunction observed in ME/CFS patients.
[...]
To summarize, some serotypes of EVs exhibit CNS tropism and have the ability to produce persistent viral infections that result in atypical and distinct chronic clinical outcomes.
[...]
Discussion

Multiple aspects of the ME/CFS pathophysiology, especially related to autonomic dysfunction, are reminiscent of chronic neurotropic enterovirus-related diseases and clinical outcomes. This fact, in conjunction with the enterovirus-like seasonality of ME/CFS epidemics, often occurring in spatiotemporal incidence with known poliomyelitis epidemics of the time, gives strong justification for the conclusion that enteroviruses have been etiological agents in ME/CFS outbreaks.
[...]
Our review emphasizes that EV-related ME/CFS literature indicates that some patients exhibit chronic enteroviral infection. Furthermore, our review highlights a number of experimental weaknesses (cohort size, tissue type interrogated, methodological approach, etc.) that exist across the EV ME/CFS literature for studies both supporting or opposing increased EV infection prevalence in ME/CFS patients vs. healthy controls. Those studies that do not support an increased prevalence of EV infections in ME/CFS patient cohorts using RT-PCR are especially confounded with issues related to incomprehensive RT-PCR primer design.
[...]
Indeed, the majority of studies interrogating muscle tissue and all studies we have identified interrogating brain tissue or cerebrospinal fluid via PCR or tissue culture have found detectable signs of EV infection.
[...]
Moving forward, studies aimed at identifying chronic EV infections in ME/CFS patients need to consider quality and types of samples to interrogate as well as methodological approaches to employ. The key samples suggested to interrogate further would include brain tissue, cerebrospinal fluid, and muscle biopsy samples.
 

Pyrrhus

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And here are the more technical bits for more technical readers:

Background Regarding Enteroviruses
[...]
Upon cellular entry, translation occurs following ribosome binding onto a type I internal ribosome entry site (IRES) located within the 5′[Untranslated Region (UTR)] of the viral genome. [...] During active infection the ratio of positive to negative strands is roughly 100:1, whereas chronic infections display a ratio closer to 1:1. [...] The 5′UTR of [enteroviruses (EV)] contains a cloverleaf secondary structure, [...] as well as an internal ribosome entry site (IRES). [...] The viral encoded RNA polymerase is error-prone due to lack of a proof-reading mechanism, resulting in high mutation rates throughout enteroviral evolution.
[...]
Steady-state infections are characterized by all cells in culture having low levels of non-lytic viral replication. [...] To date, multiple studies have shown a subset of enterovirus serotypes, including coxsackieviruses and echoviruses, are able to produce low replicative steady-state infections without cytopathic effect. This phenomenon may be caused by a number of factors including but not limited to 5′UTR terminal deletions that lead to replication deficiencies or reduced type I interferon response elicitation, faulty virion capsid formation due to incomplete capsid polypeptide processing, and alternative EV RNA mutations that lead to abnormalities such as stable and atypical double-stranded RNA complex formation that inhibits further viral positive strand synthesis. In the context of ME/CFS, 5′UTR terminal deletions and/or atypical dsRNA complex formation are notable, as they have been shown to occur in a proportion of ME/CFS patient cohorts in multiple studies. [...] Low levels of viral replication result in EV RNA levels so small that they may be past the lower limit of detection.
[...]
Detection of Enteroviruses
[...]
Across the enterovirus and virus literature at large, a number of methodologies are used to detect the presence of enteroviral infection in patients. In the early years of virus detection, biological approaches such as serological testing and cell culture methods were employed. [...] The main disadvantages to cell culture are that inoculation depends on quality of the patient sample and requires variable and sometimes extended time periods to allow detection. Some enteroviruses, especially persistent enterovirus variants, do not produce [Cytopathic Effect (CPE)] in cell culture. Without CPE, screening for viral nucleic acid or protein would be necessary.

Serological testing is confounded by several factors. First, enteroviruses often produce clinical disease before the appearance of antibodies, making their detection retrospective. Furthermore, enteroviruses and rhinoviruses have extensive antigenic heterogeneity and lack cross-reacting antigens, so that many different antigens would be needed to detect anti-EV antibodies. [...] Commercial labs with serological tests for EVs are far from comprehensive. For instance, the Enterovirus [...] kits sold via Virotech Diagnostics detects 14 [...] of the roughly 120 known EV serotypes. The Enterovirus Antibody Panel lab test provided by ARUP Laboratories similarly detects 14 EV serotypes [...] although the serotypes differ slightly. Negative detection of EVs via these commercially available serological tests does not conclusively eliminate the possibility of an EV infection.
[...]
The most popular detection method for identification of enteroviruses is RT-PCR, with amplification directed at conserved regions of the enterovirus genome, including those encoding the 5′UTR, 3Dpol and VP1. VP1 is the region of choice to conduct enterovirus typing. However, low sequence similarity amidst the approximately 120 enterovirus serotypes means that no one primer set is robustly comprehensive so that RT-PCR methods would have a lower chance of identifying novel EV serotypes than unbiased sequencing. RT-PCR experiments that use primers directed at the 5′UTR of enteroviruses can be problematic if the enterovirus contains mutations within the primer binding region, as is known to happen during persistent infection. Traditional RT-PCR approaches have reduced ability to identify novel enteroviruses that could be etiological agents in new diseases.
[...]
To date, ME/CFS studies reporting the use of tissue culture for EV detection have used [Cerebrospinal Fluid (CSF)] and feces in 1 and 4 studies, respectively. The singular CSF study reported two EV infections in a cohort of 4 patients, while the 4 fecal studies reported an increased EV infection prevalence in 2 of 4 studies, with cohorts ranging from a 22–25% prevalence across patient cohorts. [...] Although the prevalence of EV infections in these studies was generally shown to be significantly increased compared to healthy control cohorts, limitations in patient sample types and cell culture models may have led to findings that underrepresent the prevalence of EV infections in patient cohorts. Of the five cell culture studies, one study used only one cell type, 3 studies used two cell types and one study used three cell types. [...] Furthermore, the investigators were searching for [Cytopathic Effect (CPE)], and EVs present in chronic infections commonly undergo genetic changes which reduce CPE. An example of the inadequacy of CPE is a report that inoculated cell cultures were negative for CPE production in human fetal lung fibroblast and tertiary monkey kidney cell cultures but were nevertheless positive upon RT-PCR. [...] Studies reporting the absence of enterovirus infections in ME/CFS patient cohorts using tissue culture approaches had small sample sizes and incomprehensive cell culture systems. Small sample sizes along with the fact that EVs harboring 5′UTR deletions do not produce CPE means that no definitive conclusion can be made about the absence of EVs from the data in these studies. Furthermore, fecal samples usually identify only acute enterovirus infections and not chronic ones that might be in secondary infection sites.
[...]
Studies between the 1970s and late 1990s that screened for EV infections in ME/CFS patients largely focused on serological testing. The diversity of testing employed in a total of 20 serological-based ME/CFS studies included neutralization, complement fixation, micro-metabolic inhibition, ELISA, indirect immunofluorescence, and VP1 antigen detection tests. In total, 16 of the 20 studies found an increased prevalence of [Coxsackievirus B (CVB)] signals in ME/CFS cohorts with positive findings ranging from 8 to 90% compared to the positive findings in healthy control cohorts that ranged from 0 to 65%. The vast majority of studies evaluated the presence of antibodies directed only against CVB enteroviruses, with a few exceptions. [...] A notable study was performed in 1997, in which neutralization tests for 11 enteroviruses [...] found that 100 out of 200 tested patients had elevated enteroviral titers. Although serological testing in ME/CFS cohorts generally shows an increase in the prevalence of EV antibodies, the findings often lack clinical specificity as a high prevalence of EV antibodies are found in the general population from previous exposure. In a retrospective study, it cannot be known whether the enterovirus infection occurred before or after ME/CFS disease onset without having paired sera from both time periods.
[...]
The enteroviral capsid protein VP1 is commonly used for [immunohistochemical] identification of enteroviral virions in ME/CFS patient tissues. In total, 5 studies have used this technique on a variety of patient sample types, including muscle, gastrointestinal, and brain tissue. Of these, 4 out of 5 studies identified the presence of VP1 capsid proteins in patient tissue. The muscle tissue study did not detect VP1 staining in samples of a cohort of 30 ME/CFS patients, despite RT-PCR signals that indicated the presence of EV RNA in 13 of the same 30 patients. The authors suggested that the difference in PCR and VP1 immunochemistry resulted from persistent but latent enteroviral infection in patient muscle tissues, in which no detectable amount of virion particles were being produced. The remaining 4 studies showed positive VP1 staining in both gastrointestinal and brain tissues. Gastrointestinal samples exhibited positive staining rate of 82% in two patient cohorts. [...] The ME/CFS [cohort] showed dsRNA staining for 64% [...] of patients. [...] Because persistent/chronic EV infections with reduced CPE and viral replication typically have a 1:1 ratio between enteroviral positive and negative RNA strands, finding a high rate of dsRNA in patient tissues indicates the likely presence of persistent enteroviral infections.
[...]
The 4 Northern blot studies used muscle tissue biopsies and were all positive for viral RNA, indicating an EV prevalence between 21 and 50% in ME/CFS with control cohorts showing a prevalence between 0 and 1%. The two RNAseq studies were negative for the presence of EV in blood, whether or not blood was taken before or after an exercise stress that exacerbated subject symptoms. While RNAseq is a more comprehensive approach to enterovirus detection than Northern blots, these studies cannot be directly compared since one used muscle tissue and the other assayed blood samples. With regard to EV studies that applied RT-PCR methods, 5 of the 17 reports indicated no significant difference in EV prevalence between ME/CFS and control cohorts. [...] A list of all 8 PCR approaches/methods, indicating the primer sets employed in RT-PCR experiments, was first compiled, and then each PCR set was examined for its effectiveness for detection of all 117 known EV serotypes. [...] As mentioned earlier, EVs are known to exhibit mutations in the 5′UTR that result in replication deficiencies. Interestingly, all 8 PCR methodologies used primer pairs targeting the 5′UTR with the exception of method 5. [...] This is an important consideration as patients infected with EV variants exhibiting 5′UTR deletions may not be successfully targeted by the primer sets employed across these PCR methodologies. In conclusion, PCR studies aimed at identifying EVs in ME/CFS have been crippled by the use of incomprehensive primer sets that target potentially deleted portions of the viral genome.
I must say that I am a bit disappointed that they did not mention the enterovirus viroporin, which I feel is a critical concept that might help explain a persistent, steady-state enteroviral infection of the nervous system...
 

bread.

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And here are the more technical bits for more technical readers:



I must say that I am a bit disappointed that they did not mention the enterovirus viroporin, which I feel is a critical concept that might help explain a persistent, steady-state enteroviral infection of the nervous system...
Hi,

Could you please give me a rundown on what you would like to see being tested on me/cfs patients in relation to enteroviruses in a research setting?

I am going to privately fund some researchers (can not say who at this point in time) to do research into the possible connection of Enteroviruses and me/cfs and I am looking for an independent opinion.

Thank you.
 

Pyrrhus

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Hi,

Could you please give me a rundown on what you would like to see being tested on me/cfs patients in relation to enteroviruses in a research setting?

I am going to privately fund some researchers (can not say who at this point in time) to do research into the possible connection of Enteroviruses and me/cfs and I am looking for an independent opinion.

Thank you.
Personally, I would like to see quality studies that look for markers of any RNA virus (not just enterovirus) in lymph node biopsies from the lymph nodes at the back of the neck in ME patients. (with a control group, of course)

Why lymph nodes at the back of the neck? This is where the "garbage" from the brain is washed out to. If we want to know what is going on in the brain without opening up the skull, we will have to look
through these "garbage bins" of the brain.

I don't think that muscle biopsies are going to be reliable tissues to investigate, since animal studies suggest that persistent enteroviral infection of muscle tissue tends to be a focal infection, not a diffuse infection. This means that the enterovirus occupies a few small pockets of the muscle tissue, not the entire muscle tissue. A muscle biopsy could easily miss these few small pockets of enterovirus.

Looking in cerebrospinal fluid (CSF) is easier than looking at lymph node biopsies, but it won't tell you as much about what is going on inside the brain since CSF largely flows into the brain, not out of the brain:

1626579225776.png


What kinds of testing could be performed on these lymph node biopsies? Ideally, one would perform RNA-seq to identify any and all viral RNA in an unbiased fashion, and quantify the relative amount of positive strands and negative strands.

In addition, there could be immunohistochemical testing for the enteroviral VP1 and 3Dpol proteins, and for dsRNA. Whereas in an acute enteroviral infection one would expect high VP1 and 3Dpol with low dsRNA, a persistent enteroviral infection would probably show low VP1 with high 3Dpol and high dsRNA. It might also be a good idea to stain for prominent flaviviral and coronaviral proteins as well, since flaviviruses and coronaviruses seem to use similar techniques as enteroviruses to persist in the nervous system.

I hope this helps.
 
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bread.

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Personally, I would like to see quality studies that look for markers of any RNA virus (not just enterovirus) in lymph node biopsies from the lymph nodes at the back of the neck in ME patients.

Why lymph nodes at the back of the neck? This is where the "garbage" from the brain is washed out to. If we want to know what is going on in the brain without opening up the skull, we will have to look
through these "garbage bins" of the brain.

I don't think that muscle biopsies are going to be reliable tissues to investigate, since studies have found that persistent enteroviral infection of muscle tissue tends to be a focal infection, not a diffuse infection. This means that the enterovirus occupies a few small pockets of the muscle tissue, not the entire muscle tissue. A muscle biopsy could easily miss these few small pockets of enterovirus.

Looking in cerebrospinal fluid (CSF) is easier than looking at lymph node biopsies, but it won't tell you as much about what is going on inside the brain since CSF largely flows into the brain, not out of the brain:

View attachment 43975

What kinds of testing should be performed on these lymph node biopsies? Ideally, one would perform RNA-seq to identify any and all viral RNA in an unbiased fashion, and quantify the relative amount of positive strands and negative strands.

In addition, there should be immunohistochemical testing for the VP1 and 3Dpol viral proteins, and for dsRNA. Whereas in an acute enteroviral infection one would expect high VP1 and 3Dpol with low dsRNA, a persistent enteroviral infection would probably show low VP1 with high 3Dpol and high dsRNA.

I hope this helps.
Thank you, where are these lymph nodes in the neck - to be more exactly if you can - or will it be obvious to researchers and doctors alike?

Is this a standard procedure lets say in lymphoma?
 

Pyrrhus

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Thank you, where are these lymph nodes in the neck - to be more exactly if you can - or will it be obvious to researchers and doctors alike?
That's a great question. It has only been a few years since we have begun to map out the lymphatic system of the brain, so we still don't know exactly which lymph nodes connect to which lymphatic pathways. In addition, different people can have very different anatomical locations of lymph nodes.

Therefore, the locations of the lymph nodes may have to be determined on a per-patient basis, possibly by using an intelligent application of PET tracers. Another technique might be to subject the patient to a large sustained cognitive exertion, wait 24 hours, and then look for any enlarged lymph nodes.

Is this a standard procedure lets say in lymphoma?
I'm not sure. It's probably standard procedure for cancer, but not in other situations.
 

Oliver3

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This is not scientific at all but just an anecdote.
I have a raised lymph nodes which corresponds with catching suspected covid.
I had CFS previous to this.
Practicing the buteyko method with long breath holds got rid of the lymph nodes the first time and I experienced the most horrific " cleansing reaction".
The second time, with long covid, the first time I took quercetin, the PTSD feeling vanished and the lymph node went down albeit this time, only for an evening.
Just thought I'd share
 

sometexan84

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Hi,

Could you please give me a rundown on what you would like to see being tested on me/cfs patients in relation to enteroviruses in a research setting?

I am going to privately fund some researchers (can not say who at this point in time) to do research into the possible connection of Enteroviruses and me/cfs and I am looking for an independent opinion.

Thank you.
What about an in vitro study regarding Pegylated Interferon Lambda for persistent Enterovirus B infection, with and without an antiviral?

Feel free to PM me
 

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It is interesting to note that the timing of the first recorded outbreaks of ME happen to correspond with the emergence of the new virus Enterovirus A71. A coincidence, perhaps...

Enterovirus A71 evolved from Coxsackie A16 at some point between 1929-1952, so I would consider Coxsackie A16 to potentially be a serious persistent infection:

Evolutionary Genetics of Human Enterovirus 71: Origin, Population Dynamics, Natural Selection, and Seasonal Periodicity of the VP1 Gene
https://journals.asm.org/doi/full/10.1128/JVI.01019-09
 

sometexan84

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"What about an in vitro study regarding Pegylated Interferon Lambda for persistent Enterovirus B infection, with and without an antiviral?"

@bread. Funding limitations are the only reason why a study like this has not begun. If you have resources for this, please look into this, consider it. It's literally a matter of acquiring around $30,000.

Clinical trials for this drug can't begin until a study like this is completed.
 

sometexan84

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@Martin aka paused||M.E. @Oliver3 @lint7
I thought of something like this initially... I guess I just assumed reaching $30k worth of donations was not realistic.

Math

So, if these stats hold true:

1) Avg gofundme donation is $66
2) 1 in 3 donate more than once

...and let's assume someone donating for a 2nd time donates less that 2nd time, let's say $33 (or half their initial donation).

That would be $33 + $66 = $99 (let's round up to $100). So 1 in 3 donate $100. And the rest donate $66.

If I did my math right, this comes to $77.56 per donor (not donation).

That's 387 people needed to contribute in order to reach $30,000. 387 just does not sound feasible.

If we're more motivated than others...

But, let's assume we are all more motivated than others, and so our donations are larger than the gofundme averages.

Instead of $77.56, let's make the crazy assumption of $100 per donor.

That's still 300 people needed in order to reach $30,000. And this is still assuming some of the 300 make multiple donations.

300 would be incredibly difficult to pull off. So many things would have to go right. And we'd have to dumb it all down so people understand the what and why of the study. We'd have to really figure out how to make it resonate, and really really "sell" it. And even then, there'd still be a lot of grunt work or leg work to promote awareness of the campaign to multiple groups of people on multiple channels and platforms.

Of course, there's also the possibility of trying to get the budget lowered from $30k.

Anyway, thoughts?