Enterovirus PCR

Springbok1988

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Hello all,

Has anyone, after getting a high ARUP Enterovirus result, gotten a serum Enterovirus PCR? Or gone a step further and gotten a muscle or stomach biopsy tested?
I got had high antibody titers for Coxsackievirus B1, B2, and B5 on the ARUP test. Because of that I requested a serum PCR and am waiting for the results. I’m unsure if I should push for a muscle or stomach biopsy if it is negative.
Thank you for your input!
 

Hip

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Has anyone, after getting a high ARUP Enterovirus result, gotten a serum Enterovirus PCR? Or gone a step further and gotten a muscle or stomach biopsy tested?
I got had high antibody titers for Coxsackievirus B1, B2, and B5 on the ARUP test. Because of that I requested a serum PCR and am waiting for the results. I’m unsure if I should push for a muscle or stomach biopsy if it is negative.
Thank you for your input!
Were your antibody titers equal to or higher than the 1:160 threshold that Dr Chia uses to indicate chronic active infection?


Unless you have severe bedbound ME/CFS, your blood PCR for enterovirus will usually be negative. This is because in ME/CFS, the infections are in the tissues, but there is not much found in the blood. The high antibody levels often detected in ME/CFS are likely a reflection of the infection in the tissues.

For enterovirus ME/CFS, Dr Chia found that even using the most highly sensitive PCR testing kit, he was only able to get positive blood PCR in around 12% of the milder ME/CFS cases. See here.

If you test your muscles or stomach though, PCR is more likely to be positive, as those are the areas where the infection is found.


Dr Chia uses oxymatrine as an enterovirus ME/CFS treatment, among other therapies.
 

Springbok1988

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Were your antibody titers equal to or higher than the 1:160 threshold that Dr Chia uses to indicate chronic active infection?
Yes, for CVB2 my titer was 1:320.
My doctor and I decided to do a serum PCR test first, even though I expect it to be negative, because it is much less invasive.
This infectious disease doctor was unaware of Enterovirus induced post viral fatigue. So I am the one leading the way. I’m just wanting some reassurance that I should request a biopsy if the serum PCR is negative.
 

Hip

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I’m just wanting some reassurance that I should request a biopsy if the serum PCR is negative.
I am not entirely sure what Dr Chia does in terms of testing, but it is possible he may consider elevated antibody titers sufficient evidence to treat with his enterovirus therapies.

Stomach biopsy can be useful if you were negative for enterovirus by antibody testing, as the biopsy can detect enteroviruses which are outside of the range detected in the ARUP antibody tests. Sometimes people are negative by antibody testing because they may have an enterovirus ARUP cannot detect. ARUP only test for 5 echoviruses, but there are a total of 32 known human echoviruses.
 

Hip

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This infectious disease doctor was unaware of Enterovirus induced post viral fatigue.
Unfortunately in the world of ME/CFS, there is very little in terms definitive facts or proof. So we say that viruses such as the enteroviruses and herpesviruses "have been linked to ME/CFS" or "are associated with ME/CFS". But at this point we cannot say for sure that the viruses are the cause of this disease.

This is true of many other diseases as well. For example, multiple sclerosis is strongly linked to EBV, but we still do not know for sure whether EBV is the cause of MS.


So when high antibody levels are seen, ME/CFS doctors often work on the assumption that these viruses are playing a causal role in your illness, and may treat the viral infection with antivirals or immune-boosting drugs or supplements. And sometimes this gets good results. But it is still an assumption that viruses are the cause.

So this may be why your ID doctor was not aware of the enterovirus link to ME/CFS, as it is not yet a proven cause of ME/CFS.
 
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Springbok1988

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I’m honestly not sure he had ever even heard of post viral fatigue or ME at all, other than long COVID. I haven’t met a doctor who knows anything about CFS other than the name. I may be biased because I was studying virology/vaccinology for my masters when my fatigue worsened and I had to drop out. But my symptoms did get much worse after a coxsackievirus-like illness. And if there is evidence of an active infection, I’d like to find it. But if others with high enterovirus antibodies have tried and failed, it may not be worth it. I’ve just read so many papers linking to two and saying that they have found active infections.
 

Hip

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I may be biased because I was studying virology/vaccinology for my masters when my fatigue worsened and I had to drop out.
That's very interesting, it would be good to get your perspective on the non-cytolytic coxsackievirus B and echovirus infections found in the tissues of ME/CFS patients.

During the acute infection in the host, enterovirus actually undergoes a tiny mutation in its genome, and this genomic change converts regular enterovirus into non-cytolytic enterovirus, which is in effect is a different virus with a different lifecycle. Non-cytolytic enterovirus does not follow the normal lytic cycle of regular enterovirus, but instead lives inside cells as naked viral ssRNA and dsRNA.

These intracellular non-cytolytic enterovirus infections are slow at replicating and exist only at low levels in the tissues, but do have mechanisms whereby they can transmit and infect other cells. And they can persist for decades or indefinitely.

One signature of non-cytolytic enterovirus is the relative amounts of positive strand and negative strand viral ssRNA. In a normal lytic enterovirus infection, you find a ratio of about 100 times more positive strand than negative strand. But in non-cytolytic infection, the ratio is closer to 1:1.



In Dr Chia's $250 own lab test for detecting non-cytolytic enterovirus in stomach tissue sample, he normally uses the enterovirus VP1 protein stain to detect enterovirus. In his research papers though, he uses both the VP1 stain and PCR testing. The stain seems to have more sensitivity than PCR: his paper found 82% of ME/CFS patients' stomach tissues positive by the stain, but only 37% by PCR.

British ME/CFS researchers in the 1990s used to use skeletal muscle biopsies of ME/CFS patients to detect enterovirus by PCR. These British biopsy studies are listed in this MEpedia article. But muscle biopsies are painful and leave a scar, so that's why Dr Chia pioneered a new approach using stomach biopsies instead, which are not painful.

With biopsies, you can also get sampling error, as you may resect a piece of muscle tissue which happens not to contain any virus. So then you can get a false negative. For his stomach biopsies, Dr Chia asks the gastroenterologist to cut a sample of stomach tissue in an inflamed area of the stomach, so that you are more likely to snip a piece of tissue containing the virus.



I don't have any background in biology or medical science, but I got interested in these non-cytolytic enterovirus infections, and over a 10 year period, slowly learnt a few things about them.

I wrote this MEpedia article on non-cytolytic enterovirus (though it has never been checked by a virologist, so may contain errors).

Non-cytolytic enterovirus is not only found in ME/CFS, but is also found in (and may be the cause of) type 1 diabetes, dilated cardiomyopathy, and several other chronic diseases.
 
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sometexan84

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I have high B2, B3, B4, and Echovirus 11 titers, via ARUP labs. I'm actually waiting on the PCR results, but like Hip said, it'll prob be negative.

I believe the ARUP titers are enough though. If I need to get a biopsy in order to prove the infection to a doc, I'll do it.

Also, you want to retest. I had B2 titers that started at 1:40, then climbed to 1:160, then to 1:320. I won't be comfortable til they ALL come back at or below 1:10.

There are others here who have found the virus w/ a stomach biopsy.
 

Pyrrhus

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I apologize in advance for being overly pedantic... :rolleyes:

During the acute infection in the host, enterovirus actually undergoes a tiny mutation in its genome, and this genomic change converts regular enterovirus into non-cytolytic enterovirus
Technically speaking, it is the infection that changes from a cytolytic infection to a non-cytolytic infection.
Technically speaking, a virus by itself is neither cytopathic nor non-cytopathic- it is the infection caused by the virus that is. I apologize again for being pedantic, I can't help it!

And in the case of enteroviruses, it seems likely that it is not only the gradual loss of the first 40 base-pairs of the 5' end that is responsible for the conversion of the infection from cytopathic to non-cytopathic. It seems likely to be a combination of a number of changes, both in the virus itself as well as in the infected cell that are responsible for this conversion. As always, more research is needed to disentangle these contributing factors!

One signature of non-cytolytic enterovirus is the relative amounts of positive strand and negative strand viral ssRNA. In a normal lytic enterovirus infection, you find a ratio of about 100 times more positive strand than negative strand. But in non-cytolytic infection, the ratio is closer to 1:1.
Yes, finding an equal amount of positive and negative strand viral RNA is the most direct way to determine whether or not an infection has "converted" into a non-cytopathic infection. But just to clarify- the reason why equal amounts of positive and negative strand RNA is found is because the virus appears to have been trapped in its dsRNA intermediate state. Of course, to quantify the amount of positive and negative strand RNA in an assay, one must first denature the dsRNA into ssRNA. I apologize yet again for being pedantic, I really can't help it! :headslap:
 

Hip

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It seems likely to be a combination of a number of changes, both in the virus itself as well as in the infected cell that are responsible for this conversion.
Yes, that is indeed the case.

Prof Nora Chapman, who discovered the mechanism by which regular enterovirus converts into non-cytolytic enterovirus within host cells, says that this conversion can only take place in non-dividing cells (quiescent cells), like muscle cells. You will not get a non-cytolytic infection forming in rapidly dividing cells, like liver cells. Rapidly dividing cells are immune to non-cytolytic enterovirus infection.

Prof Chapman figured out that the reason you need a quiescent cell to create a non-cytolytic infection is that in quiescent cells, a cellular factor called hnRNP C is not available to the virus. This factor is very important for efficient enteroviral replication. Once enteroviral replication in the cell is greatly hindered due to a lack of hnRNP C, conditions in the cell become ripe for a non-cytolytic infection to emerge.

The MEpedia article section: "How does lytic enterovirus transmute into the non-cytolytic form?" gives more details.



I was wondering whether there might be drug which could to induce hnRNP C, in order to try to clear the non-cytolytic infection, by converting the infection back to a regular lytic infection. But unfortunately I could not find any hnRNP C inducers.
 

sometexan84

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I still think Peginterferon Lambda is absolutely the best bet for eradicating enterovirus. There are already studies and trials showing how effective it is against lots of other enteric viruses, similar to coxsackie and echovirus.

And aside from one research article showing how Interferon Type III (Lambda) has a massive antiviral effect on Coxsackie (as well as demonstrating how it evades the natural interferon response), there's been no talk about Peginterferon Lambda for persistent Coxsackie or Enterovirus B infections.

If I can't convince Eiger Pharmaceuticals to grant me off-label or compassionate use for their lambda drug, I'm going to try and convince them to do a small trial for use in persistent enterovirus b infections. They're already in Phase 3, which should make this at least possible.