Since the first association of the retrovirus XMRV with chronic fatigue syndrome in 2009 in the US, subsequent studies have failed to detect evidence of infection in patients from the US, Europe, and China. These studies were potentially compromised by a number of factors, such as differences in patient characterization, geographic locations, clinical samples used, and methods used to detect the virus. These and other potentially confounding conditions have been addressed in the most comprehensive study to date on the association of XMRV with CFS.
In the introduction to their paper, published in the Journal of Virology, the authors note other problems with many of the studies of XMRV in CFS patients:
- Too small control populations
- Patient and control samples collected at different times
- Investigators generally not blinded to sample identity
- PCR assays that rely on conservation of viral sequence mainly used
- Limits of detection, reproducibility, and precision of assays unknown
- Controls for each step that would identify analysis not done
- Insufficient numbers of negative controls included
- No study included positive samples from the original 2009 patient cohort of Lombardi et al.
To address these issues, the authors collected blood from 105 CFS patients and 200 healthy volunteers in the Salt Lake City area. One hundred of the patients fulfilled both the CDC-Fukuda and the Canadian consensus criteria for diagnosis of ME/CFS. The patients were selected from a clinic that specializes in the diagnosis and management of CFS and fibromyalgia.
New blood samples were also collected (by a third party) from 14 patients from the original study by Lombardi et al. The samples were blinded for subsequent study. Detection of viral nucleic acids was done using four different PCR assays. Anti-XMRV antibodies in patient sera were detected by ELISA. Finally, virus growth from clinical specimens was attempted in cell culture. The authors used the multiple experimental approaches reported by Lombardi and colleagues.
Let’s go through the results of each assay separately.
PCR for viral nucleic acids. Four different quantitative PCR assays were developed that detect different regions of the viral genome. The assay for pol sequences has been used by several groups and is the most specific PCR assay for XMRV. Three other PCR assays were also used that target the LTR, gag and env regions of XMRV DNA. These assays could detect at least 5 viral copies of XMRV DNA. The precision and reproducibility of the PCR assays, as well as their specificity for XMRV, were also demonstrated. DNA prepared from white blood cells of 100 CFS patients and 200 controls were negative for XMRV. For every 96 PCR reactions, 12 water controls were included; these were always negative for XMRV DNA.
XMRV antibodies in human sera. To detect XMRV antibodies in human serum, a portion of the viral envelope protein, called SU, was expressed in cells and purified from the cell culture medium. The SU protein was attached to plastic supports, and human serum was added. Any anti-XMRV antibodies in human sera will attach to the SU protein and can subsequently be detected by a colorimetric assay (we have discussed this type of assay previously). This assay revealed no differences in the amount of bound human antibodies for sera from CFS patients or healthy controls. Some of the patient sera were also used in western blot analysis. Recombinant XMRV SU protein was fractionated by gel electrophoresis. The protein on the gel is then transferred to a membrane which is mixed with human serum. If there are anti-XMRV antibodies in the human serum, they will react with the SU protein on the membrane, and can be detected by a colorimetric assay. When rabbit anti-XMRV serum was used in this assay, the SU protein was readily detected. None of the human sera analyzed by this method were found to contain antibodies that detect SU protein.
Infectious XMRV in human plasma. It has been suggested that the most sensitive method for detecting XMRV in patients is to inoculate cultured cells with clinical material and look for evidence of XMRV replication. The XMRV-susceptible cell line LNCaP was therefore infected with 0.1 ml of plasma from 31 patients and 34 healthy volunteers; negative and positive controls were also included. Viral replication was measured by western blot analysis and quantitative PCR. No viral protein or DNA was detected in any culture after incubation for up to 6 weeks.
Analysis of previously XMRV-positive samples. Blood was drawn from twenty-five patients who had tested positive for XMRV as reported by Lombardi et al. These samples were all found to be negative for XMRV DNA and antibodies by the PCR and ELISA assays described above. In addition, no infectious XMRV could be cultured from these 25 samples.
Presence of mouse DNA. After not finding XMRV using qPCR, serological, and viral culture assays, the authors used the nested PCR assay described by Lo et al. Although positives were observed, they were not consistent between different assays. This led the authors to look for contamination in their PCR reagents. After examination of each component, they found that two different versions of Taq polymerase, the enzyme used in PCR assays, contained trace amounts of mouse DNA.
Given the care with which these numerous assays were developed and conducted, it is possible to conclude with great certainty that the patient samples examined in this study do not contain XMRV DNA or antibodies to the virus. It’s not clear why the 14 patients resampled from the original Lombardi et al. study were negative for XMRV in this new study. The authors suggest one possibility: presence of “trace amounts of mouse DNA in the Taq polymerase enzymes used in these previous studies”. I believe that it is important to determine the source of XMRV in samples that have been previously tested positive for viral nucleic acid or antibodies. Without this information, questions about the involvement of XMRV in CFS will continue to linger in the minds of many non-scientists.
At the end of the manuscript the authors state their conclusion from this study:
Given the lack of evidence for XMRV or XMRV-like viruses in our cohort of CFS patients, as well as the lack of these viruses in a set of patients previously tested positive, we feel that that XMRV is not associated with CFS. We are forced to conclude that prescribing antiretroviral agents to CFS patients is insufficiently justified and potentially dangerous.
They also note that there is “still a wealth of prior data to encourage further research into the involvement of other infectious agents in CFS, and these efforts must continue.”
Clifford H. Shin, Lucinda Bateman, Robert Schlaberg, Ashley M. Bunker, Christopher J. Leonard, Ronald W. Hughen, Alan R. Light, Kathleen C. Light, & Ila R. Singh1* (2011). Absence of XMRV and other MLV-related viruses in patients with Chronic Fatigue Syndrome. Journal of Virology : 10.1128/JVI.00693-11
I agree. I would even go further and say that rather than donating directly to ME research, we should mostly donate to effective ME advocacy orgs that will get us the proper funding from the government since the chance is only slight that our meager donations to research alone will solve this disease. I hope and assume ANIDA will be this org. Time will tell.
Just a small amendment, it should say “In Samples WPI Previously Declared to be Positive”. Whether or not they were actually positive is unclear, that is the whole point.
Roy I am afraid you are talking out of your bottom……:
“And when the original WPI results are clearly and completely invalidated in the next few months, as I now expect they will be”.
XMRV/HGRV/MLV is here to stay that is a fact. If you think its going to be over in the next few months you havent been following the science or story.
Please offfer up your hypothosis for the cause of ME. Its your opinion to offer.
There you go again talking as if the WPI is a lone voice. Why is it that you and your chums push this lie over and over again.
I am interested in Alter, the CC, the NIH and the WPI. They all offer the strongest hypothesis of ME causation if you actually follow the issue. they are yest to be disproved regardless of what you say.
Please can you tell us about the PACE trial and the science behind that also as RRM seems to be refusing to engage in that one.
I essentially agree (though to quibble, an error in online supporting material is less grave than an error in the paper). It’s possible that WPI, NCI and CC made errors in Lombardi et al.
Unfortunately, your definition of “unvalidated assays” seems to be any assay which does not detect that which you assume must be there. Your assumption may be incorrect. Any good scientist would at least be willing in this case to question the original assumption.
Using your approach, geocentrism would still dominate because Copernicus, Kepler et al used unvalidated methods. And the fact that the Flat Earth Society still exists is just proof that a number of people will never concede the truth, no matter how overwhelming the evidence.
Guest – I like the cut of your jib!
Why are you changing the subject, we are not talking about where the virus came from. You already know this virus. HIV assays that are developed after virus was discovered didn’t all use clinical samples for positive control. Besides a clone is a wild type virus if the sequence is the same. Who are these regulators? With all these claims, you are actually accusing the reviewers and editors of the Journal of Virology with publishing sloppy experiments.
A single cause for a social consrtuct is a misnomer Johan. I am not saying that all other leads should not be followed.
The point is about assataining who has a HGRV from the ME/CFS population and treating those people accordingly. You cant just say “let us move on”. That makes no sense. Its like saying move on from theories of various cancers because different people may have different causes for their cancer. This would be especially dangerous in any illness where you had identified a group of people between 70-80% with a possible shared underlying cause.
HGRV is the best hypothesis for causation of ME as a neuro immune disease with multi coinfections and other bio markers like spinal fluid issues etc. These other issues are not causation hypotheses just as co infections etc in aids are not the cause of HIV.
The whole point is to identify a sub group from a social construct, “CFS” which enables you to treat them accordingly.
HGRV XMRV is in its infancy dropping it now would be like dropping HIV in 1983 and concentrating on pneumonia for the illness AIDS.
There has to be something was was missed. I tested positive by serology and have been taking truvada and Raltegrarvir and feel so much better. I am no longer bedridden. Imy lower back pain has reduced drastically although some brain fog still exists. Other issues still exist but 50% of them have disappeared and 25% have had relief. More research has to be done but saying not to take antiretrovirals would have left me incapacitated with a resting heart rate of 110 which is now down to 85. Night sweats are gone and my raynauds symptoms in my fingers has disappeared. Other issues have cleared up so it just can’t be a coincidence.
But you are missing the point RRM. The above named have validated the original findings. Something you keep ommiting when you try to convince people that the WPI is a lone voice. If the Alter study etc had been a 0/0 it would have proved nothing.
That is the very point. 0/0 studies cannot claim to prove anything either way why do you keep pretending it is scientific to claim they have proved something. The weight of evidence against 0/0 studies is that the assay and the method is inadequate.
Tell us about the PACE trial and how it was another scientific study on the issue of ME RRM!!
I can’t believe that some people liked that comment. 10 years? Are you kidding me? You openly contradicted yourself even in your own comment.
“It was 10 years after the discovery that they isolated HIV”
AIDS was discovered in 1981, virus was isolated in 1983. Maybe that’s 10 dog years, but in humans that’s 2 years. The first drug as you said was available in 1987, so it’s 4 years AFTER the isolation of the virus!
I’d say we should do both 😉 Certainly we should all sign up with those orgs, so they can show they have a large constituency and will be able to mobilize many people for actions.
Given the often mentioned prevalence of ME/CFS of 0.25% our donations are not meager at all, a couple of millions of people can raise incredible sums, even if most are poor.
We need to get it from everywhere.
“XMRV/HGRV/MLV is here to stay that is a fact.” Clearly there is no point in further discussion of this issue. When you state a hypothesis as a given “fact”, and therefore something that cannot be invalidated no matter what the future findings or data, there is no point in going on. You’ve already made it clear that any study that does not support the WPI findings is flawed by definition.
I hope an objective qualitative study can be achieved in the future. All I know is that whenever I’ve been on a course of antibiotics(rarely), I have had short term relief from symptoms. That’s gotta mean something in someone’s medical journal !!
Yes, i agree. But the conclusion is only correct if the methods in this latest study are as effective as the ones used by Lombardi et al. and Lo et al. I can’t judge this.
I also agree that now i’m expecting a good enough reaction from the WPI.
But that XMRV can’t be found in the blood would not at all rule out there is an association with ME/CFS. If the 4% in healthy controls are correct and i’m waiting for Dr. Singh’s reaction on this (not personal of course, but i’m sure she will have to adress that), then basically this latest study merely answered the question what methods should not be used, because it didn’t pick up XMRV in the controls (and also in at least some ME/CFS cases who should also have a similar rate, i guess).
Then we would need to find out where XMRV can best be found in the body and with what methodology and then look in ME/CFS patients in this way.
It does not have to be an undetectable strain. It could be exactly the same strain as in Schlaberg et al.’s prostate cancer patients and/or healthy controls, as long as the assays used in this latest study have not shown they can detect those in the blood. Or not?
Quote “XMRV/HGRV/MLV is here to stay that is a fact.”
This frame of mind is exactly what I am afraid of. If Lipkin doesn’t find anything, if let say Ila Singh doesn’t find XMRV in prostate cancer anymore, … XMRV will still continue to exist if only in the minds of a group of patients seeing conspiracy theories everywhere, going the way of the anti-vaccine movement, alien abductions, chem-trails, … That statement “… here to stay is a fact” has nothing to do with science or facts, it’s a belief.
The original paper on XMRV and CFS was a demonstration of science in action, but so are the negative papers (including the one of Singh and the contamination papers). If XMRV turns out to be just another dead end, then at least Mikovits can take credit for getting a lot of scientists interested in ME/CFS. Don’t ruin this by alienating/bullying/attacking those scientists. We might need them.
Tecala, I am not questioning your personal experience in this matter. But what you describe is also almost a textbook example of a placebo effect, especially the short-term relief from symptoms.
What does the PACE trial have to do with anything here?
With move on I meant letting go of the obsession with XMRV, not completely dropping it. It looked very promising, but it is becoming increasingly unlikely to be the (single) cause of ME/CFS.
There is no such thing as HGRV, it’s still just a hypothesis. No more than wishful thinking.
There are no primers or conditions mentioned for Nested PCR in Lombardi et al. There are for PCR as performed on 11 samples at the Cleveland clinic and there are for Nested RT-PCR on gag.
There is no point in I asking you to quote the section that list primers or conditions for Nested PCR as they do not exist.
This is the sort of nonsense we have to deal with on a constant basis. Notice the overstretching headline, unsupported by the underlying science:
Chronic Fatigue Syndrome Is Not Related to XMRV Retrovirus, Comprehensive Study Finds
http://healthcare.utah.edu/publicaffairs/news/current/05411Singh.html
Until she came up negative the Singh study was expected to be that objective qualitative study. The next one is Lipkin, but if he comes up negative it will not considered objective either (at least by some).
The short term relief you experience can (it is a possibility, not a certainty) be explained by the placebo effect.
The placebo effect can also explain the patients who got (a bit) better on anti-retrovirals. In one of her posts Jamie Deckoff-Jones mentions briefly patients who got worse on anti-retrovirals. Unfortunately those patients are probably not in a position to blog.
What would make you happy? Changing the word “Finds” to “Concludes”? Or how about “Comprehensive Study Concludes That XMRV Retrovirus Is Not Present In CFS Patients”?
Lombardi et al
“we isolated nucleic acids from PBMCs and assayed the samples for XMRV gag sequences by nested polymerase chain reaction (PCR) (5, 6). Of the 101 CFS samples analyzed, 68 (67%) contained XMRV gag sequence. ”
“Nested RT-PCR for gag sequences was done as described (5) with
modifications. GAG-O-R primer was used for 1st strand synthesis; cycle
conditions were 52oC annealing, for 35 cycles. For second round PCR,
annealing was at 54oC for 35 cycles.”
Mikovtis et al
“In our experience from performing the
multiple methods on the same 57 blood
samples(…) nested RT-PCR
of plasma nucleic acid or PCR from cDNA
from unactivated PBMCs”
Switzer et al (2010)
“The XMRV specific assay uses the primers GAG-O-F and GAG-O-R and GAG-I-F and GAG-I-R for the primary and nested PCRs, respectively, and conditions as previously described [11,12]. This is the same nested PCR test used by Urisman et al. and Lombardi et al. to detect 413-bp XMRV gag sequences in prostate cancer and CFS patients, respectively [11,12].”
van Kuppeveld
“Primers were as described, except for the reverse primer of the nested reaction (GAG-I-R), which was replaced by GAG-I-R2 to yield a 92 base pairs reaction product (we used primer GAG-I-R2 because it produced less background in the nested reverse transcription polymerase chain reaction). ”
RRM: “it is very much in the interest of the government and the CDC to solve the ME/CFS problem….Government bodies want to solve this problem, the scientists involved want it…”
It is not necessary to resort to conspiracy theories, exactly, to understand why the CDC and the NIH have dropped the ball on this illness since the 1980s, and only now, very belatedly, seem to be picking it up again. Unfortunately, science does not always live up to its own ideals of disinterestedly seeking the truth, and public agencies operate like other large institutions; often in their own interests or according to the individual prejudices of their leadership, rather than in the interests of those they are supposed to serve. This is not conspiracy theory: it’s a fact of how the real world often works.
Look at the economic impact figures of the disease burden of ME/CFS (estimates range from 10 billion to 20 billion dollars per annum in the US in lost productivity, disability payments, medical costs etc) and then look at the amount of money allocated by the NIH for ME/CFS research for the current fiscal year: $6 million. That is not a typo. HIV/AIDS gets $3 billion a year.
The CDC estimates there are between 1-4 million people in the US with ME/CFS. (And the wide range in that figure points very clearly to a major part of the problem: the CDC has failed to adopt a rigorous enough definition of the disease to even be able to tell the public whether an “extra” 3 million people *actually* have the disease. Such a definition – much more rigorous and careful – is available in the Canadian Consensus definition published by Carruthers et al a good eight years ago, but the US has inexplicably failed to adopt it. David Tuller wrote a very good piece for the New York Times a few months ago explaining why rigiorous disease definitions matter to properly studying a disease.)
Let’s say for the sake of argument that with a more rigorous definition we might be looking at 1 million people with more rigorously (CCC) defined ME/CFS in the United States. Let’s further guesstimate that 25% of those are ill enough to be completely disabled from working; the rest have various degrees of debility that impact their quality of life, economic productiveness, etc.
Looking for current statistics in the US on HIV/AIDS, I find that the estimates are that 1 million in the US are currently HIV+ and that just under half a million are living with AIDS. I don’t see with any quick Googling how many people are currently disabled within those numbers (the SSA does not consider a person with HIV or AIDS to automatically be “disabled,” the determination will be based on ability to work, since medication regimens have become so good that many with HIV can go on a long time without developing AIDS or ARC and without significant detriment to their ability to work.) HIV has gone from a death sentence to a serious but manageable chronic disease. An incredible, wonderful achievement that took a lot of serious effort by a lot of researchers, and a lot of serious financial commitment. But that effort didn’t get underway in the beginning based merely on the seriousness of the disease and its economic impact: there had to be a lot of hard-fighting advocacy in the early years.
It’s not necessary to know about Stephen Strauss and the things he said about ME/CFS patients; the scanty to non-existent evidence with which ME/CFS was declared at a certain point to be “probably pyschosomatic” via official NIH press release; the fumbling and bumbling of the CDC in trying to investigate outbreaks of ME/CFS in the 1980s, and the politics which choked off research funding and even diverted research funding earmarked for ME/CFS into other pet projects at the CDC (this isn’t conspiracy theory; there was a 1998-2000 GAO investigation that confirmed this had happened, to the tune of $8.5 million) …or any of that, to get a very clear picture of the seriousness with which US public health agencies are investigating ME/CFS, All you have to do is look at the NIH budget for the current FY to see where priorities have been placed, and compare the level of funding on ME/CFS to a range of other conditions…like seasonal allergies…or rare diseases that affect only a few hundred.
A vast conspiracy to hide the obvious truth? Not exactly. It has never been obvious what causes ME/CFS, although there are lots of objective findings showing biological abnormalities in this patient group. The US health agencies, by the way, don’t yet accept any of these objective findings as diagnostic. You can be unable to get out of bed, and still denied disability benefits because your illness is still a matter of someone’s taking your word for it that you are sick. Plenty of the general public and PLENTY of doctors still think it’s a bogus disease, or psychosomatic in some way, or an invention of the Internet on the level of “Morgellon’s disease.”
These beliefs are not held because of rational economic calculations; they have to do with ignorance, prejudice and stigma. If you ask me, the buck absolutely stops with the public health agencies. It’s up to them to provide leadership on this disease: find a cause and hopefully a cure or at least better treatments, sure. They could also help us out by putting some muscle behind validating a diagnostic test, so that we aren’t faced with trying to “prove” what is currently unprovable by their guidelines. They could change the stupid, belittling, and misleading name of “Chronic Fatigue Syndrome.” (That wouldn’t even cost much money.) They could give a much clearer and stronger message to the press about the seriousness of the disease, which could help a little bit with the distorted public opinion of this disease and the stigma attached to it.
They could, in short, pick up the damn ball that they dropped 25 years ago and fulfill their duty to the public. Sheer economics, as you say, would seem to make this a sensible course of action. I also happen to think it’s their moral duty. Let’s see if we get solid dollars to back up recent promises of more focused and productive action.
Where is the evidence that any of those assays work?
Scientists question data by testing it.
Whether there is a god is not the discussion here. The data in Lombardi et al can be tested.
Do you understand why lowering the stringency of the primers is important. If you cannot answer that question then you are failing to understand the issue.
You see this is the whole point RRM. If Switzer or anyone else can find the HGRV XMRV at all in any small amount of patients it is further proof of a new HGRV and kills of the contamination theory.
It also shows that a 0/0 study is using methods incapable of detecting XMRV as they are not even able to detect population levels of XMRV.
The existence of the HGRV, XMRV is not in question except by conatmination theorists like yourself.
The blood studies are to ascertain the levels found in people with ME not to prove XMRV actually exists. If Singh sampled some 300 patients in her blood study and found zero positives it shows her assay on blood to be incapable.
Zero detection = incapable assay.
Primarily, because self-report is unreliable, and a method such as the one you suggest does nothing to screen out confounding factors.
Surely you’ve been following the long deconstruction of the PACE trial results where we have discussed in great detail the problems with self-report and how such reports can be influenced by various factors. In the case of the PACE trial, a very clear and expicit bias in what the different treatment arms were *told to expect* from the particular treatment they were receiving; in the case of Internet groups, the self-reinforcing groupthink that rewards wishful thinking.
“Today I feel better, the drugs must be working!” “Today I feel worse, I must be detoxing,” (followed by chorus of supportive messages: “Stick with it, you’re so brave, right on for you!”)
They did as the regulators would have demanded they have proof that the assays work.
How do we know the virus? What do you think is being tested for? That is the subject you raised.
A clone is not clinically positive samples.
“Study does not find XMRV in ME Patients’ Blood.”
In light of the fact that XMRV was shown to appear only transiently in the blood of infected monkeys, you can say neither that this study is comprehensive (it did not look at tissue) nor flatly that ME is not related to XMRV.
Please explain the PACE study to us and the government bodies behind it. Tell us how the scientists in the PACE study wanted to resolve the ME issue and how watertight the study was.
Was the PACE study “shitty” as you described the book Justin refers to or was it a genuine attempt at science and reporting the truth to the media.
RRM do you even know what PACE is and are you a patient of ME. What is your interest in this issue?
Your arguments are becoming more and more ridiculous by the minute. They are so transparent and people are waking up to you and questioning what your purpose is in this debate.
The more you talk the more the uniformed will see the holes in your logic.
Thankyou for setting the record straight Mindy. It is amazing that so called scientists and a Professer can claim the opposite here as fact and glaringly mislead people.
One would think people “in the know” would be more informed and check their facts.
But then this is only a matter of life and death for some of us.
Firstly, the IAP assay has not been validated, but Lo has tested samples with this and found no contamination. Perhaps when the assay is validated they will publish this data. However, it is also not known if the IAP assay can also identify human IAP sequences or those from mice transferred by the virus.
Second, Lo and Lombardi et al ruled this out by testing for mouse contamination, none was detected. The CDC has also ruled it out in 20 samples from the WPI.
1. Who are “they”?
2. I didn’t raise that subject. I just said you don’t need positive clinical samples as positive control and even though I repeated it this 4 times now, you still don’t comprehend it.
3. So first you said a clone is not wild type virus, and now you are saying a clone is not clinically positive. I never said it was, because you don’t need clinically positive samples.
4. I never denied the existence of this virus. We know it because we isolated it.
1. Who are “they”?
2. I didn’t raise that subject. I just said you don’t need positive clinical samples as positive control and even though I repeated it this 4 times now, you still don’t comprehend it.
3. So first you said a clone is not wild type virus, and now you are saying a clone is not clinically positive. I never said it was, because you don’t need clinically positive samples.
4. I never denied the existence of this virus. We know it because we isolated it.
To clarify Singh found 22% of her prostate cancer patients to be infected with XMRV which is a new Human Gamma Retrovirus.
Not finding it in the blood doesnt mean it is not there in the blood and certainly does not mean it is not present in the patient at all.
That is science 101.
So you are bowing out of a scientific discussion on the subject of XMRV a HGRV in a huff because you have nothing credible to offer to the basic notion of the scientific method.
Wow, If you cant grasp the starting point it maybe is a good decision on your behalf to stop posting naively on it.
Can you tell us anything about the PACE study instead and how scientific it was and what it proves?
Enlighten us how the scientists involved came to their conclusions and convinced the mass media of their findings.
Johan why do you think people are still fighting the Psychogenic explanation of ME after decades of it being proven wrong.
Is it because the “scientists” behind this explanation are wright and the patients are wrong and clinging to unscientific beliefs.
Science is science not a tally of papers.
Be very wary of an “ultimate” decision.
No it is not;
“Correction: I meant to say they obtained 25 names of CFS patients from
WPI, not 25 samples. The 25 individuals were then bled, but only 14
had been previously positive.”
These WPI samples were freshly drawn. And we know from the macaque study, that XMRV is found transiently in the blood.
My goodness if this is your level of understanding and is the way you want events to turn God help us all whatever Hypothesis is ever developed for ME.
One minute you claim a HGRV is wishful thinking then you say it is unlikely to be the single cause.
Somewhat contradictory statements. For those who may be infected are they not entitled to be treated if it is the cause of their own disease state.
In 1983 would you have made the remark in the early days of HIV to “let go of the obsession with HIV”.
What an astonishing statement you make above.
I don’t claim to own your preferred hypothesis of ME but do tell what it is in your opinion.
Further more the idea that “XMRV is the single cause of ME” is a definition based argument. It is better to ascertain of those who may be positive to a new HGRV what treatment do they need for their given illness.
The reason I am here debating XMRV is because that is why this thread exists. So saying to move on from it is a little daft.
Anyone who doesn’t want to discuss it is not forced to come here where the whole purpose is to discuss it.
Well I am asking you if well funded studies can be critically analyzed for claiming things that are simply not true.
You seem to think that is not the case.
So tell us do you think the PACE trial adds any value to the ME argument. There are numerous studies and claims by the proponents of the PACE trial that CBT and GET can cure, treat or reduce symptoms of ME. Its simple do you agree with them?
If not why not, there is lots of “evidence” to prove it is the case.
Nested in their paper is used as short for Nested RT-PCR. The only mention of Nested PCR is for conditions for sequencing full-length XMRV genomes. The 67% was based on Nested RT-PCR. See the other post where others support that it was Nested RT-PCR. Or how about this zany idea, read the paper.
The positive sample from the WPI was not used in the study. He used 22Rv1
“As a positive control for the polymerase chain reaction assay, we used nucleic acid isolated from 22Rv1, a prostate carcinoma cell line (American Type Culture Collection number CRL-2505) that was recently shown to contain multiple integrated copies of XMRV and to produce high levels of infectious virus.”
Since in any case it seems that finding XMRV in blood is near impossible, why don’t we quit looking for it in the blood?
The experiments injecting XMRV into monkeys have shown that XMRV is quickly cleared from the blood and have also shown which tissues are XMRV reservoir in the long term. So why don’t we look for XMRV in biopsies from these tissues, and use these results to do epidemiology studies to determine:
– the prevalence of XMRV in the population
– the potential association between XMRV and various diseases, CFS, prostate cancer, and others?
Aren’t there banks of biospied tissues available in many institutions that would allow even preliminary studies to be undertaken? One could even use fresh biopsies from patients suffering from various conditions requiring one, for the simple purpose of obtaining data on general population prevalence of XMRV infection?
Because the fact remains that XMRV does exits at least in lab cultures, and that it is infectious. So in any case it would be of prime interest for Science to know whether of not it is present in the population and wether or not it is associated with disease.
RRM where do you get your ideas from?
If 14 patients repeatedly test positive at different times this is further evidence that the WPI methods are validated. If they didn’t test positive repeatedly this would weaken the WPI argument as one might claim contamination was the cause of an initial positive test.
You claim that certain groups cant find their own samples positive consistently (which is not true) then when they offer Singh consistent positives she shows further evidence that her assay is in capable by testing the most scrutinised samples negative.
Not only this, in the whole study she shows her blood assay is incapable of detecting population levels of XMRV at the very same time that Switzer declares XMRV to be present in 1.9% of prostate samples he tested.
Switzer did not claim contamination he decalred it XMRV.
You will not get passed the regulators without proof.
You actually don’t help yourself here. A clone is not clinically positive samples. Do you understand the meaning of the last three words?
Where was the virus isolated from?
Lombardi samples were validated.
You don’t say who the quote is from, but it is irrelevant because they are wrong. You cannot claim an assay works without proof. The study was negative.
Ah I see, we are going in a placebo effect direction now. Laughable!
What next CBT and GET cures ME?
Come on guys what do you make of the PACE trial I know some of you are itching to say.
Who are these “regulators” you keep bringing up?
My intention is not to “help myself”
Just because you isolated a virus from the blood of a person, doesn’t mean that you need to use that sample for validating your assays in later studies.