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CROI (Retrovirology and Opportunistic Infections, Boston) on XMRV and CFS

Cort

Phoenix Rising Founder
Cort wrote:

Dusty Miller approached this community in a way that was unethical: he hadn't received IRB(Institutional Review Board) approval of his protocol.

He wanted to use an untested assay that, as I understand it, would falsely indicate contamination when people were actually XMRV+, undermining the work of the WPI. This is a very important issue. Until things are much clearer people should be very chary about participating in experiments which are using this assay. As I recall, he wanted to perform a "definitive" study. Don't they all.

Miller claimed to have a great familiarity with the political issues animating the M.E. world. If he did and still was going to enroll patients from Buchwald's clinic, which views the disease as factitious, then he was not truthful.

If his commitment to the XMRV project could be swayed just by virtue of his conversations with other retrovirologists what does that say about his earlier assertion that he was on board with the association of XMRV and M.E.?

His participation on me/cfs forums was not as innocent as you describe; he gave as well as took. If a handful of voices, some abrasive perhaps, could sway this scientist from significant research, what sort of scientist is he?

Miller ultimately agreed to use both the mtDNA and IAP assays in the study - that is my understanding and I think you missed some points that both Ecoclimber and I raised; a) Ecoclimber said they missed a deadline and b) my understanding was that as he talked to more retrovirologists Miller's interest faded. Dusty Miller came on those Forums for at least a month, I would guess - he hung in there through thick and thin - it wasn't the abrasiveness of the patients that ultimately dissuaded him - it was other things.

I would note that he is still engaged in XMRV research and he is apparently collaborating with Dr. Silverman on XMRV integration. He did do work with nerve cells and XMRV and my understanding is that that paper will be published. So he is still working on XMRV - he's just not, at least at this moment, doing the patient study.

I think the ME/CFS Forums were ultimately quite helpful in an important way....We thought that going to Miller with his reputation was enough but the uproar prompted us to contact Dr. Mikovits -and doing that surely made the study stronger - since he was able to incorporate her concerns into it.

I didn't say that Miller was going to use Buchwalds clinic. Please read what I said carefully. He had no intention of doing that -that was why he went straight to the online community. He was actually trying to AVOID doing that. The uproar, however, could have pushed him in that direction. He was accused of lying, of having no experience with gamma retroviruses, warnings were sent across the internet that he was out to get XMRV.... - someone filed an ethics complaint against him. As Ecoclimber said, institutions don't like that kind of publicity.

The IRB! What an issue. The ethics complaint was that he approached patients too quickly ( he didn't Ecoclimber did and Miller protested that)...therefore the community felt he shouldn't be trying to find XMRV.....which I felt was putting the horse behind the cart..Deciding not to back a study partially because you're concerned about IRB issues is missing the mark IMHO.

Usually good studies are derailed because of picky IRB Boards and researcher rue how long it takes to get through them. (I know one that just was denied by a picky IRV Board). Now you have patients actually going to the IRB board and saying stop this study....that's a head turner in my opinion. I wonder if that has ever happened before.
 

currer

Senior Member
Messages
1,409
Unfortunately, the unbalanced way in which some doctors (or researchers), who get involved with CFS/ME are treated by patients, convinces them that we have a primary psychological disorder.

My consultant did not believe this and was very sympathetic to his patients. But once the orchestrated attacks on him commenced, fuelled by internet-spread misinformation, I could see that he was having second thoughts, despite himself.

I do not know how to quell this misguided paranoia on the part of some people. I suppose it will only cease once ME/CFS is accepted as a valid illness. and that is more in the hands of the politicians than doctors or researchers. Because I think the ultimate cause for the paranoia is the political injustice with which we have been treated. It only helps "them" if we argue with and attack those who try to help us, even if their approach is not perfect. So what, why should it be!
 

akrasia

Senior Member
Messages
215
Cort wrote:

I didn't say that Miller was going to use Buchwalds clinic. Please read what I said carefully. He had no intention of doing that -that was why he went straight to the online community. He was actually trying to AVOID doing that. The uproar, however, could have pushed him in that direction. He was accused of lying, of having no experience with gamma retroviruses, warnings were sent across the internet that he was out to get XMRV.... - someone filed an ethics complaint against him. As Ecoclimber said, institutions don't like that kind of publicity.

Cort, it is my terrible fate to be condemned to read what you write carefully. And that's not the impression you gave. The impression I got was that Miller rebuffed, in a fit of pique, was going to enlist patients for his work from Buchwald's dismal clinic.


The IRB! What an issue. The ethics complaint was that he approached patients too quickly ( he didn't Ecoclimber did and Miller protested that)...therefore the community felt he shouldn't be trying to find XMRV.....which I felt was putting the horse behind the cart..Deciding not to back a study partially because you're concerned about IRB issues is missing the mark IMHO.


IRBs are important for many reasons, the primary one in this case is the ability of a participant to give informed consent. No one should participate in work whose ramifications might compromise their health or the hope of finally receiving effective, appropriate treatment for M.E. This is quite a serious ethical issue, not just on a personal level but on a collective one as well.
 

thegodofpleasure

Player in a Greek Tragedy
Messages
207
Location
Matlock, Derbyshire, Uk
I think the ME/CFS Forums were ultimately quite helpful in an important way....We thought that going to Miller with his reputation was enough but the uproar prompted us to contact Dr. Mikovits -and doing that surely made the study stronger - since he was able to incorporate her concerns into it.

"Uproar" is thus a very necessary part of the scientific process.
It is the patient community's response to what is happening in the scientific world AND SHOULD NOT BE CENSORED

Equally, playing the part of "Sugar Daddy" is not a free pass to avoiding scrutiny or criticism.

True philanthropists, who want to remain anonymous, don't tend to talk about their philanthropic generosity and certainly don't participate in forums such as this !
 

Marco

Grrrrrrr!
Messages
2,386
Location
Near Cognac, France
I've seen Ecoclimber's name crop up on threads on this and the other forum although to my knowledge I have not contributed to them.

I have no axe to grind but find this whole discussion puzzling. While any assistance is to be welcomed, particularly from someone who is also ill, the stakes are high and people are understandably concerned that any future XMRV research does not make the same perceived (there will be two sides to any argument) mistakes as with previous studies.

My concern is not with believing or not in XMRV but in the association being research thoroughly and fairly and in accelerating research into all biomedical aspects of ME/CFS.

I have a few questions for Ecoclimber if he is still around. They are honestly posed and I'm insinuating nothing.

Firstly, I find it strange that he should post here with currently unsubstantiated information about XMRV possibly being a recombinant XMRV contaminant. While this may be true (the information), and may in due course be shown to be true, what advanatage did it offer the people on this forum to be aware of this in advance?

You state that you only became aware of ME/CFS a year ago and that, as a philanthropist, you wished to help. Why the interest in CFS as it would be known in the states? Given the public perception of CFS as merely 'fatigue' I can think of many other poorly understood illnesses that would on the face of it be more worthy of financial support?

You state that you represent or have close ties with the Gates Foundation. The Gates Foundation is deeply involved in vaccination programmes worldwide including HIV/AIDS and Bill Gates has made some pretty strong statements lately against the 'anti-vax brigade'. You will not find any anti-vax people on this forum although you will find much concern that vaccines safety should be taken very seriously and concerns investigated thoroughy (as someone nicely put it lately - 'just because I support the recall of Toyotas with brake problems doesn't make me anti car').

However you will find people here who have been subject to accusations of potentially contributing to the development of ARV drug resistance through the unlicensed/unofficial trialling of HIV ARVs in response to being tested as XMRV positive. More generally this point has been used to argue against even the clinically controlled trialling of ARVs which could be a major factor in proving causation. More generally still, I have been concerned that even if XMRV is proven to be the causative pathogen in ME/CFS, clinical decisions will need to be taken on the costs v benefits of ARV treatment for a population widely considered to be afflicted only with 'fatigue'. ARV drug resistance will also factor into these decisions.

You may not be in a position to answer, but do you believe that the Gates Foundation would support the treatment of ME/CFS with ARVs if and when that was proven to be an effective therapy?


Given the above and the recent experiences with trying to get involved in XMRV research, there are many other areas of biomedical ME/CFS research that are crying out for financial support downstream of actually identifying the pathogen if XMRV is too much of a 'hot potato'. What would prevent you and/or the Gates foundation getting involved here?
 

anciendaze

Senior Member
Messages
1,841
While I'm still trying to get information on the actual paper, I see a post of mine on the other forum has become part of the debate here. I'm trying to avoid ad hominem arguments, and I'm assuming ecoclimber is honestly reporting what he has been told. Scientific conferences are rumor mills, and the placement of this presentation near the end pretty well limits the amount of critical attention it can receive.

As for my "specious argument", the counter argument assumes PCR will detect viral sequences even when they have been hypermutated. We know of one family of enzymes actually doing this. We don't know all causes. We have seen that replication-competent virus can be cultured from hypermutated sequences. We have seen PCR fail in many unrelated cases.

One cell in 100,000 sounds more definitive than it is. Millions of cells are involved.

In the end this comes down to assumptions, a chance coincidence of sequences is very unlikely. The path from the research with nude mice to the R22v1 cell line is obscure. The path from XMRV in a xenograft to mutated XMRV which reverts to infecting murine cells is simple and natural. Sequence divergence is more common than convergence. PCR failures are well known in unrelated research.

If the virus is in human prostate cancer cells, contamination of both nude mice and R22v1 is predictable. This will also explain antibody response which sequence contamination will not.

If you start with the certainty this cannot be in humans, you must assume a contamination theory, no matter how convoluted, explains everything. The idea that these endogenous sequences just happened to be present at the same time in the same mice, and not in other mice, is a long shot before you throw in the probability of a recombination event producing a replication-competent species being observed by chance in the lab.
 

cigana

Senior Member
Messages
1,095
Location
UK
Dear Ecoclimber,

I want to add my name to the list of people who would dearly love it if you would reconsider. I didn't know your history or work backstage, thanks for explaining and thankyou deeply for everything you've done so far.

The funny thing is, while all of this is going on on this forum (and the other one), all of the people I know at my local support group don't even know these forums exist. They don't really know anything about XMRV either. They are all just a bunch of normal people who got really ill one day and are trying to push through.

Here in the UK some people have suffered so bad I am not exaggerating when I compare it to a holocaust. One day it will come all come out. It seems like with every other disease, they have research and lucky breaks. We have never had a lucky break. Instead we have some psychiatrists who kick us while we're down, they control the research grants, the media and treatment. So no matter what a handful of people have said about you, I think you could go down in history as the person who stops a holocaust. One might even say you have a moral imperative! But I don't want to put so much pressure on you to scare you off. Just to let you know the thanks you'll receive from millions should make all this forum nonsense seem insignificant!

Maybe just pull out of XMRV. Do you know of Dr Kerr's work in the UK? He announced on national TV that he should be able to get treatment or even a cure in about a year based on his genetic findings. Then he lost his job. A relatively small amount of money there could go such a long way for millions of desperate people.

Cheers,

Mark

p.s. Here is that video of Dr Kerr on TV:
[video]http://www.investinme.org/Multimedia/Meridian-Taylor/Meridian%20ME%20Experiences%20150kbps.wmv[/video]
and for a reminder of how bad things are here, this is Sophia Mirza on UK TV:
[video]http://www.investinme.org/Multimedia/Meridian-Wilson2/Converted/MM100-768KbpsDSLorLanorCab.wmv[/video]
 

SilverbladeTE

Senior Member
Messages
3,043
Location
Somewhere near Glasgow, Scotland
Cigana
people think that "evil" men who condemn millions to suffering and death have ot be monster, ghouls, "touched by the dark gods", some sign about them that they are foul or insane...
alas, no.

The utter banality, the stupidity, the selfish stupidity and impenetrable hubris of evil people is...alas, unremarkable in general, until you really see what they do, what they cause, how uncaring they are of their victims, often stating they are doing it "for the greater good!", and it often takes too much hard work for ordinary people to comprehend.

Tobacco companies set up a secret project to enrich themselves, to "protect" their product, you can go read about this if wish.
SO they made their cigarettes more addictive and toxic, so harder to break the habit and faster to get addicted.
Researchers were bribed, bought off, or attacked, ridiculed depending on their stances
you name it, all kinds of crap.
The end result is 100 MILLION dead by the end of the 20th century, somewhat more than were killed in the entire horrors of World War 2!
By end of this century the death toll will be one BILLION.

Why has the SAS or SEALS not slaughtered their boardrooms? Why have we not dropped tactical nukes or MOABs on their headquarters? Why have we not napalmed and cluster bombed their facilities?
because it makes MONEY, because they pay a lot of TAX and bribes, that's why.

I have known 1 person killed by terrorism yet we slaughter those bastards, quite rightly.
I've known quite a few murdered (lost count after four by time I was 25 years old), police/courts rightly hammer those scumbags.
I have known oh maybe 20 dead by alcoholism...do the booze firms suffer, no.
Smoking has probably killed about 40+ folk I've known over the years it's hard to tell really. My mum's on an oxygen machine lot of the time because of smoking. See any jsutice for that? No.

The sons of bitches responsible for our deliberate disenfranchisment, abuse, suffering, DEATHS, are just another lot of arrogant bean-counting ass wipe scumbags who keep their hands clean and pretend, even to themselves some are surely utterly deluded, that they are doing it all "for the greater good!"
nothing remarkable about them. they lack the courage or the wisdom to be anything else.
And for that, they are going to rot in Hell, and Hell is much much worse than the traditional and erroneous "fiery inferno" concept.
Alas it's hard to feel pity for them as I should, but suffering ME is jack compared to what they've got coming and that I do feel sympathy for, even if they've brought it on themselves, it's still horrendous :/

"For those who have no mercy, there will be no mercy!"
 

cigana

Senior Member
Messages
1,095
Location
UK
Silverblade, when this is all over we'll need you to be the court's expert witness:D
 

SpecialK82

Ohio, USA
Messages
993
Location
Ohio, USA
Finally...

WOW.

Aren't we all supposed to be be pretty ill and want our live's back?

What won't help is anyone being obnoxious or beligerent or intolerant. I don't read the other forums because the people who stopped posting here and went there had a different standard of acceptable forum behaviour. Let's debate 'til the cows come home but keep it civil. If you don't keep it civil you scare people off.

I'm glad this forum seems to prioritise civility.

Whose side am I on ? Every CFS patient's side. I want my damn life back and I don't need anyone pushing that day further away because they can't make the effort to treat their fellow patients or anyone else for that matter with a basic level of respect and politeness.


FINALLY - A voice of reason. My sentiments exactly, and probably many more who read the forum but are too ill to expend energy on nonsense. I am on science side as well. Of course we want the correct diagnosis not the easiest or the fastest - the wrong one won't make anybody well.

We are all ill and frustrated and sometimes do not handle stress well, and watching all the infighting recently is heartbreaking. We are tearing each other apart for what purpose? Is feeling right more important than getting well?

We need to do everything we can to support real science and let the chips fall where they may.
 

Rita

Senior Member
Messages
235
Session 43-Themed Discussion
TD: XMRV: New Findings and Controversies
Wednesday, 1-2 pm; Room 302-304
Paper # 217
Presence of XMRV Sequences in B Cells Are Restricted by APOBEC
Jorge Carrillo1, J Blanco1, E Garcia1, J Arreal2, B Clotet1, and C Cabrera1
1Fndn irsiCaixa, Barcelona, Spain and 2Hosp Univ Germans Trias i Pujol, Barcelona, Spain

Background: A link between xenotropic murine leukemia virus-related virus (XMRV) infection and different human diseases, such as chronic fatigue syndrome (CFS) and prostate cancer, has been recently established. Given that this retrovirus can infect different tissues and cell types both in vivo and in vitro, including cells from the immune system, the identification of the cellular compartment(s) where XMRV can establish a reservoir may be useful to understand the pathology associated with this virus.

Methods: In order to determine the presence of XMRV in B lymphocytes, we have screened EBV-transformed B-cell lines available in our laboratory. The origin of these cell lines include CFS patients (n = 11; fulfilling both Fukuda and Canadian criteria), HIV+ individuals (n = 4), prostate cancer patients (n = 1), and healthy donors (n = 5). DNA was extracted from cellular dry pellets and several XMRV genes were amplified by either real-time PCR (pol) or nested PCR (gag and env).

Results: We detected 7 positive samples from 14 individuals tested by RT-PCR (4 CFS, 2 donors, and 1 HIV+ individual). env amplification identified 4 positive samples out of 21 individuals tested (3 CFS-affected individuals and 1 healthy donor), whereas gag amplification showed only 3 positive samples (1 CFS-affected individual, 1 healthy donor, and one HIV+ patient). To confirm the presence of XMRV we performed sequence analyses of gag and env amplicons. The 3 gag sequences available showed a 100% homology with XMRV gag published sequences, including the XMRV characteristic 24nt deletion which is not found in any known exogenous murine leukemia virus. Furthermore, env sequences were also homologous to previously described XMRV sequences, showing none or low variability. Interestingly, most of the observed changes corresponded to multiple G to A mutations that were accumulated in 1 positive sample, resulting in a truncated env protein and suggesting that APOBEC-related restrictions operate in vivo during XMRV infection.

Conclusions: Despite the discrepancies observed in the different PCR approaches, our data provide evidence that EBV-transformed B-cell lines harbor XMRV-specific sequences, although the establishment of this infection could be modulated by the innate restriction factor APOBEC 3G . Our data suggest that, in vivo, B cells may represent a reservoir for XMRV contributing to its potential pathogenesis.
 

Rita

Senior Member
Messages
235
ession 58-Poster Abstracts
XMRV and GBV Virus–Host Interaction
Tuesday, 2-4 pm; Hall D
Paper # 239
Limited Evidence of XMRV Infection in Different Populations in Spain, Including Patients with CFS and PC
Miguel Arredondo*1, J Hackett2, F de Bethencourt3, A Trevio1, D Escudero4, A Collado5, P Labarga1, P Swanson2, V Soriano1, and C de Mendoza1
1Hosp Carlos III, Madrid, Spain; 2Abbott Labs, Abbott Park, IL, US; 3Hosp La Paz, Madrid, Spain; 4Hosp Germans Trias i Pujol, Barcelona, Spain; and 5Hosp Clin, Barcelona, Spain

Background: The recent discovery of xenotropic murine leukemia virus-related virus (XMRV) and its association with chronic fatigue syndrome (CFS) and prostate cancer (PC) has generated enormous interest. However, conflicting results have been obtained by different investigators. In this study, we examined populations of archived specimens from Spain for evidence of XMRV infection using serological and molecular methods.

Methods: Plasma and peripheral blood mononuclear cells (PBMC) were collected from 1103 patients: 437 CFS and/or fibromyalgia (FM), 69 PC, 149 HIV, 31 human T cell lymphotropic virus type 1 (HTLV-1), 81 chronic hepatitis B, 72 chronic hepatitis C, 18 autoimmune disease, and 246 blood donors. Plasma samples were evaluated for the antibodies to XMRV transmembrane protein using a prototype ARCHITECT XMRV p15E research assay (Abbott Diagnostics, Abbott Park, IL). DNA extracted from PBMC were tested for the presence of XMRV gag sequences using nested PCR following the Lombardi protocol. Samples that were gag PCR positive were further evaluated using nested PCR for XMRV env sequence.

Results: Of the 1103 plasma samples, 3 (0.3%) were p15E seroreactive with a sample/cutoff (S/CO) of >1.0: 2 HTLV-1 patients (S/CO 1.2 and 4.8) and 1 chronic hepatitis C patient (S/CO 1.04). Of the 662 PBMC DNA tested by PCR, 4/69 (5.8%) samples from PC patients with stage T1 (2), T2 (1), or T3 (1) cancer were repeatedly positive for XMRV gag sequences. However, XMRV env sequences could not be PCR amplified from DNA of these 4 patients. Moreover, no patients had both p15E antibodies and detectable XMRV gag sequences.


Conclusions: In this study, limited evidence of XMRV infection was identified in a large population of Spanish patients with various medical conditions including CFS and PC previously associated with XMRV infection. However, further confirmatory studies are required. Screening for the presence of gp70 antibodies using a more sensitive serological assay is in progress.
 

Rita

Senior Member
Messages
235
Home Search Abstracts View Session Email Abstract Author


Session 43-Themed Discussion
TD: XMRV: New Findings and Controversies
Wednesday, 1-2 pm; Room 302-304
Paper # 216
XMRV Induces a Nonproductive Infection in Human Lymphoid Tissue
Marta Curriu1, J Carrillo1, M Massanella1, E Garcia1, B Clotet1, C Carrato2, J Blanco1, and C Cabrera1
1Fndn irsiCaixa, Badalona, Spain and 2Hosp Univ Germans Trias i Pujol, Barcelona, Spain

Background: Xenotropic murine leukemia virus-related virus (XMRV) has been associated with prostate cancer and chronic fatigue syndrome. In humans the virus has been found in a variety of cell types, including T and B cells, and in rhesus macaques there is evidence for viral replication in lymphoid organs, suggesting that lymphocytes are a target for XMRV. Histocultures of tonsils support productive infection with various viruses, including HIV and human herpesvirus 6. In this study, ex vivo lymphoid tissue was used to investigate the pathogenic mechanisms of XMRV.

Methods: Tonsils from 2 healthy individuals undergoing tonsillectomy were collected and cultured in small pieces (2 mm3) over gelfoam soaked in RPMI medium. Tissue blocks were left uninfected or infected with XMRV stock obtained from a 22Rv1 cell supernatant. Culture medium was replaced every 3 days. After 14 days in culture, both tissues were homogenized and cells were isolated. Viral infection was evaluated at different times in the cells migrating out the tissue and at day 14 in tissue cells by PCR, analyzing viral DNA. In addition, tissue cells were immunophenotyped by flow cytometry and the presence of envelope protein (env) was analyzed by Western blot (WB) using an antibody to SFFV env that reacts with all poly- and xenotropic murine leukemia virus.

Results: Seven days post-infection cells migrating out the tissue were positive for XMRV DNA. After 14 days of culture, tissue cells were also positive, confirming that XMRV infected human tonsil tissue in the absence of exogenous stimulation. Despite the presence of XMRV DNA, infection does not seem to be productive since tissue lysates exhibited undetectable expression of XMRV env proteins by WB. Uninfected and infected tissues showed similar percentages of T and B cells. XMRV infection did not modify the percentage of CD3 (76 and 75% in XMRV+ and XMRV– tissue, respectively), CD4 (53% vs 52%), CD8 (39% vs 40%), or CD19 cells (3% vs 1%). A deeper analysis of T cell subsets showed that XMRV infection did not modify the nave/memory cell ratio, or immune activation markers, as evaluated by the expression of HLA-DR and CD38.

Conclusions: Our data show that XMRV could be integrated into the human lymphoid tissue cells although this process does not culminate in an explicit productive infection. In addition, this infection did not result in changes of T or B cells nor an immune activation, suggesting that lymphoid tissue could be a latent tissue reservoir in XMRV infection.
 

Rita

Senior Member
Messages
235
Home Search Abstracts View Session Email Abstract Author


Session 58-Poster Abstracts
XMRV and GBV Virus–Host Interaction
Tuesday, 2-4 pm; Hall D
Paper # 237
A Sensitive Real-time PCR Assay for the Detection and Quantification of XMRV
Laura Li*, M Raines, and T Robins
Quest Diagnostics Clin Trials, Valencia, CA, US



Background: Xenotropic murine leukemia virus-related virus (XMRV) was first identified in prostate tissue from prostate cancer patients. One study reported detection of XMRV in 67% of patients with chronic fatigue syndrome (CFS), as compared with 3.7% of healthy controls. Subsequently, several studies failed to detect XMRV in CFS patients, sparking controversy in the field. Therefore, a more sensitive and specific method is needed to resolve the issue. To this end, we have developed a sensitive real-time PCR assay that reliably detects XMRV from CFS patients.

Methods: XMRV RNA was extracted from CFS patients’ peripheral blood mononuclear cells (PBMC) or PBMC/LNCap cells using Trizol reagent. RNA was reverse transcribed using Superscript VILO cDNA synthesis kit. Taqman primers and probe were designed and used in the quantitative real-time PCR assay.

Results: The performance of our quantitative real-time PCR assay was determined, and then it was compared with the current XMRV detection method. Current XMRV detection methods use a nested PCR assay followed by agarose gel detection of the PCR product. These methods are labor intensive and time consuming. A sensitive real-time PCR assay was developed to detect and quantify XMRV within 2 hours. This assay could detect as few as 1 to 2 copies of XMRV RNA molecule in the reaction. In addition, the linear range of the assay was established and quantification of XMRV virus was performed to show that the assay can be used to detect and quantify XMRV RNA simultaneously. Furthermore, specificity of the assay was confirmed by the fact that it detects XMRV but not HIV, hepatitis B virus, or hepatitis C virus. Direct comparison of 1 current method with the quantitative real-time PCR assay was carried out. PBMC from CFS patients were either co-cultured with LNCap cells or not. Viral RNA was extracted and tested with both methods. Both methods could detect XMRV in the co-cultured PBMC/LNCap. However, only the quantitative real-time PCR assay could detect XMRV in non-co-cultured PBMC.

Conclusions: Based on preliminary performance data, this real-time PCR assay could be a sensitive and quantitative tool for detecting XMRV.
 

Rita

Senior Member
Messages
235
Home Search Abstracts View Session Email Abstract Author


Session 43-Themed Discussion
TD: XMRV: New Findings and Controversies
Wednesday, 1-2 pm; Room 302-304
Paper # 220
Serologic and PCR Testing of Persons with Chronic Fatigue Syndrome in the US Shows No Association with XMRV
William Switzer1, H Jia1, H Zheng1, S Tang1, R Garcia2, and B Satterfield2
1CDC, Atlanta, GA, US and 2Cooperative Diagnostics, Greenwood, SC, US

Background: In 2009, a newly discovered human retrovirus, xenotropic murine leukemia virus-related virus (XMRV), was reported in 67% of persons from the United States with chronic fatigue syndrome (CFS) by PCR detection of gag sequences. However, 5 subsequent studies in the US, Europe, and China found no evidence of XMRV or murine leukemia virus (MLV) in CFS samples from different patient populations. More recently, another group detected MLV sequences but not XMRV in 87% of US CFS patients. Thus, more work is necessary to investigate the prevalence of these murine retroviruses in persons with CFS.

Methods: We tested blood specimens from 45 CFS cases and 42 persons without CFS from more than 20 states in the US. Plasma specimens were tested for antibodies using a new Western blot assay that utilizes purified XMRV as antigen. Whole blood DNA samples were tested using 2 generic XMRV/MLV PCR tests in the polymerase region and one for gag sequences. Plasma from the CFS patients was also tested for viral RNA using a real-time PCR test that generically detects XMRV/MLV protease sequences. CFS patients were diagnosed using the 1994 research case definition and all had a minimum of 6 months of post-exertional malaise and a high degree of disability.

Results: We found no evidence of either XMRV or MLV in all 45 CFS cases and in the 42 persons without CFS using a comprehensive testing strategy.

Conclusions: Our results, combined with previous negative studies, do not suggest an association of XMRV/MLV in the majority of CFS cases across the US. Differences in patient populations may explain the discordant test results observed in each study. Further work is required to determine the prevalence of XMRV and MLV in persons with CFS and to evaluate a causal link of these viruses found in a subset of CFS cases.
 

alex3619

Senior Member
Messages
13,810
Location
Logan, Queensland, Australia
Too much research is symptom based - and that doesn't take us very far. While it is true that many diseases are effectively treated without understanding of their causes I feel research should look for a CAUSE for a disorder which appears to be acquired, and has spread so rapidly in the last thirty years.

We need people with courage to get involved, and yes, it will sort out the men from the boys! Thats reality - its not always pretty!

Hi currer, researching causes offers a real cure. Who wants to be offered aspirin instead of surgery if something is really wrong?

Also, don't discount the Amazons - most of us are women, and so most of the advocacy is by women - look at usedtobeperkytina, a true role model, even for us guys. And while she may not have ME/CFS, Hillary Johnson has done the most fantastic job as a journalist.

Bye
Alex
 

eric_s

Senior Member
Messages
1,925
Location
Switzerland/Spain (Valencia)
Thanks Rita. But the Spanish studies are nothing new, the 2 IrsiCaixa abstracts are the same like the ones from the International XMRV Workshop in September of last year and i think the one by the Madrid group was published already. So do they have anything new? I hope so. Where is the biomarker they said they have? Did they do anything between last September and now? Sorry, if this sounds hard, it's not personal.