XMRV not detected in Dutch chronic fatigue patients

26 February 2010

dutch_cfs_xmrvThe suggestion that the retrovirus XMRV is the etiologic agent of chronic fatigue syndrome (CFS) arose from a study in which the virus was found in 68 of 101 US patients. The virus was not detected in two independent studies of 186 and 170 CFS patients in the United Kingdom. A new Dutch study has also failed to reveal XMRV sequences in 32 CFS patients.

The subjects of the Dutch study were part of a 298 member cohort. All patients fulfilled the Oxford criteria for CFS and reported debilitating fatigue for at least one year. Cryopreserved peripheral blood cells taken from 32 of these individuals between 1991-92 were used for preparation of DNA. This material was then subjected to polymerase chain reaction to amplify proviral XMRV DNA. The primer sets used were the same as those employed in the US study. Under the PCR conditions used, at least 10 copies of XMRV sequences could be detected per 100,000 peripheral blood mononuclear cells. All samples from CFS patients and from controls were negative for two different XMRV genes encoding integrase and gag proteins.

The authors consider a number of reasons why their results differ from those in the initial US CFS study. They rule out (1) technical differences; (2) The possibility that the long duration of CFS in the Dutch cohort may have led to negative results, because retroviruses integrate into the genome of the host; (3) cryopreservation; and (4) differences in cohorts. They suggest that XMRV might be involved in CFS outbreaks but not in sporadic CFS:

…the peripheral blood mononuclear cells [used in the US study] were derived from patients from the outbreak of chronic fatigue syndrome at Incline village at the northern border of Lake Tahoe, United States (1984-5). This outbreak has long been thought to have been caused by a viral infection and has been associated with a number of viruses, most notably Epstein-Barr virus and human herpes virus but firm evidence for a role of viruses in this particular outbreak has never been provided. It is possible that the study of Lombardi et al has unravelled the viral cause of the chronic fatigue syndrome outbreak, but it seems unlikely that their study demonstrates a viral association for sporadic chronic fatigue syndrome cases, such as those we tested, or represents the majority of patients. Studies of XMRV in sporadic chronic fatigue syndrome cases from the United States would be of great interest.

The authors do acknowledge the small sample size used in their study, which prevents them from statistically ruling out a role for XMRV in CFS. Nevertheless they conclude that “our data cast doubt on the claim that this virus is associated with chronic fatigue syndrome in the majority of patients.”

My office neighbor here at Columbia University Medical Center is Dr. Stephen Goff, an expert on retroviruses who has begun to investigate XMRV in his laboratory. He recently gave a plenary lecture on XMRV at the Conference on Retroviruses and Opportunistic Infections in San Francisco; you can hear some of his comments at medpage today. I poked my head in his office yesterday and asked him what he thought of the story so far. His answer: everyone needs to exchange samples, and they are not doing it. I couldn’t agree more.

Frank J M van Kuppeveld, Arjan S de Jong, Kjerstin H Lanke, Gerald W Verhaegh, Willem J G Melchers, Caroline M A Swanink, Gijs Bleijenberg, Mihai G Netea, Jochem M D Galama, & Jos W M van der Meer (2010). Prevalence of xenotropic murine leukaemia virus-related virus in patients with chronic fatigue syndrome in the Netherlands: retrospective analysis of samples from an established cohort British Medical Journal : 10.1136/bmj.c1018

  • John

    I think there are several problems/inaccuracies with this paper-

    1. The statement that the samples from the Science paper came from the outbreak of CFS which occured in Incline Village in the mid-eighties is inaccurate. Dr. Mikovits has been quoted saying something to the effect of when the WPI started looking for virus, XMRV was found in the 'sickest of the sick', ie a cohort of patients that Dr. Peterson has been following for several decades and is either entirely comprised of or includes a significant number of those who became ill in the outbreak and many of whom have went on to develop gamma clonal T-cell rearrangements and b-cell lymphomas(1). Longitudinal samples taken from this group comprises Dr. Peterson's 'personal stash', and according to Dr. Mikovits it was in samples from these patients that the WPI researchers figured they had the best chance of finding something if indeed anything was to be found, and in fact it was in these samples that XMRV first started popping up.

    However this was just a first initial study. When WPI researchers started the Science study, they took randomly selected samples from a different, larger sample cohort which is comprised of patients of Dr. Peterson's colleagues around the country.(2)

    2. The 'Oxford criteria'. The study authors don't 'rule out' what has been one of the most critical factors in CFS research for the past several decades, they simply disregard it in a single sentence. The Oxford criteria has been the bane of CFS research since it's inception in 1991. It is only- not mainly, not mostly, but only used by psychiatrists and psychologists in the UK and the Netherlands who view CFS to be a primary psychiatric and/or behavorial disorder. Prevalence rates for 'CFS' using the Oxford criteria are in the neighborhood of 2.6% in the UK, with two seperate studies on prevalence rates in the US using the 1994 CDC or 'Fukuda' criteria found .24% and .42%, respectively(3). It doesn't take a mathemagician to see that the discrepancy between the two is between 6x-10x, which itself doesn't take a brainiac to see that something is very discordant between the two. However when you take out those with co-morbid psychiatric disorders from the Oxford prevalence rates, they drop down to .5%, which is very similar to the rates found using the Fukuda criteria. Basically the Oxford criteria have been a several decades long excuse for the above mentioned psychiatrists and psychologists to include patients with psychogenic fatigue states such as anxiety disorders, PTSD, depression, etc. in their studies which they purport to be on CFS, with other CFS researchers taking great care to exclude those with the above mentioned conditions from CFS studies.

    3. As noted in the WPI's last press release(4), none of the XMRV negative studies have done what WPI considers to be a critically important step- culturing the samples. In that press release WPI notes that when they did not culture samples it was necessary to look at 4 different longitudinal samples taken from the same patient in order to find a sample that had enough virus to detect using PCR.

    1. http://www.scivee.tv/node/7030

    2. http://www.wpinstitute.org/research/research_bi

    3. http://www.iacfsme.org/IssueswithCDCEmpiricalCa

    4. http://www.wpinstitute.org/news/news_current.html

  • Charlotte

    Thanks for asking Dr Goff – good to get the opinion of a top retrovirologist without an axe to grind. His comment cuts to the chase. Excellent blog!

  • eddie

    “His answer: everyone needs to exchange samples, and they are not doing it. I couldn’t agree more.”

    + 1

    Also, John (above) is correct, use of the Oxford criteria has seriously confounded and hampered efforts to pin this thing down, much to the detriment of patients.

  • ST

    Patient cohort different, PCR method different, nothing found in controls. XMRV doesn't exist in UK – are you sure? Must have been the fantastic border police we have. A replication will satisfy everyone much more then wasting money lke this.

  • Geo

    There were some new papers from the retrovirus conference posted in the last comments section on XMRV – it sounded like there was some confusion as to how best to detect XMRV. Has anyone here been keeping up with this research, and feel up to explaining whether it has any bearing on the ongoing CFS/XMRV controversy?

  • Geo

    There were some new papers from the retrovirus conference posted in the last comments section on XMRV – it sounded like there was some confusion as to how best to detect XMRV. Has anyone here been keeping up with this research, and feel up to explaining whether it has any bearing on the ongoing CFS/XMRV controversy?

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  • pwachtel

    While this result is curious it is astounding that the researchers' commentary overlooks the elephant in the corner :- patient selection using the Oxford criteria. That the issue is not even acknowledged shows a disregard for the work of those clinicians and researchers that lead the field; that have had at least some small successes in supporting the patient community.

  • aspiecelia

    You fail to mention that the Japanese who have a great deal of expertise in this disease were able to detect the virus and you information about the study which found 68 in 101 with xmrv is incorrect, you do not even mention the study which showed xmrv in >90% and those samples were from people from all over the place. The studies done in the UK had big problems. I guess this all leads me to wonder what your agenda is.

  • dancer

    Why are they not exchanging samples and how can we get them to?

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  • http://www.virology.ws profvrr

    One of the authors of the Dutch study told me
    “Concerning your remark that people should start to exchange material,
    this is exactly what I proposed to Judy Mikovitch, but I have not yet
    received an answer”.

  • CBS

    Here is a link to the actual full presentations at the CROI conference:

    http://www.retroconference.org/2010/data/files/

    All the XMRV stuff is Friday. Dr. Goff's plenary talk 'Mouse to Man' is first thing. He starts with a really clear description of what he has learned about XMRV over the last 35 years. He also talks about some clear differences in detection using different receptors as well and provides a list of possible reasons for the discordant findings and his take on each at the present.

    Also found at: http://app2.capitalreach.com/esp1204/servlet/tc

    Later (at 9:30) there are a few additional presentations on XMRV. One that I found interesting was Dr. Hackett from Abbott (last speaker in this session -it takes a while to load the video and slides but is well worth it).

    He talks about the macaque study and makes a few interesting points and raises a few questions.

    Dr. Hackett presents data that raises questions about the prevalence of XMRV in the population – perhaps less than 0.1%. If that holds up it would be hugely good news for the blood supply.

    He ends with a discussion of the challenges in detecting antibodies in XMRV and the need for amplification possibly due to issue such as viral life cycle, length of time between infection and disease, and possible impacts of XMRV itself on the ability of the immune system to mount a response.

    Also found at: http://app2.capitalreach.com/esp1204/servlet/tc

    I'm really please to see that Dr. Goff so involved in getting at what is going on with XMRV. Clearly there is a lot to learn about this bug in humans.

  • http://www.virology.ws profvrr

    We spoke twice on TWiV with Jason Rodriguez, a postdoc of Steve Goff's
    who works on XMRV. See http://www.twiv.tv/2009/09/20/twiv-50-xmrv/ and
    http://www.twiv.tv/2009/10/25/twiv-55-mice-lie-…. I
    plan to record a conversation on XMRV with Goff next week and post it
    to TWiV.

  • KTSword

    This a a science blog by a well respected virologist. There is no agenda other than good science. The blog owner has clearly stated previously that he considers CFS a devastating disease.

    Your reference to findings by the Japanese have not been published nor has the WPI, the Cleveland Clinic or the NCI published any further findings specific to XMRV in CFS.

    When such findings are published in a scientific journal I'm sure they will be noted here and elsewhere.

    I recommend reading all the posts on this blog regarding XMRV. There is much to be learned.

  • CBS

    I want to strongly second the response by KTSword. There is no agenda here. The people that Dr. Racaniello is talking with are on the forefront of trying to figure out what is going on with XMRV (in prostate cancer and CFS). Drs. Goff and Singh (former colleague at Columbia now at the Univ of Utah) amongst others are actively trying to understand what, if any, role XMRV plays in CFS.

    I am a 'CFS' patient being seen by one of the top CFS/Infectious Disease docs in the country who is presently working with Dr. Goff to investigate these questions. I have found that Virology Blog is by far the most thorough and balanced source of virology info on the web (I know in some corners that's setting the bar pretty low but this is good stuff – right now I am working my way through Virology 101). I'm learning a lot and there is a lot to learn, especially for those of us coming to this from a patient's perspective (without a strong background in virology).

    I'm really looking forward to next week's TWiV conversation with Dr. Goff.

    Hang in there.

  • CBS

    Dr. Suzanne Vernon, Medical Director of the CFIDS Assoc of Amer. recently wrote an article on the pressing need to standardize methods and share information. In what was a surprisingly public plea, Dr. Vernon addressed the authors of the Science paper, imploring them to share more information: http://www.cfids.org/xmrv/021510study.asp

    “Until methods are standardized and the scientific community is provided information about the specific characteristics of the CFS subjects (and controls) who tested positive in the Science paper, be prepared to read more negative studies. Hopefully the Science investigators will make this information available before interest in XMRV being associated with CFS fades and becomes yet another foiled attempt at solving CFS. Achieving scientific consensus on the role of XMRV in CFS certainly warrants more research and greater collaboration, as do so many other important discoveries being made.”

    In a very short time, Dr. Vernon, formerly of the CDC, has made significant strides towards creating a unique collaborative and coordianted effort across a range of scientist to more fully illuminate processes underlying CFS. See her “Association Research Program Update” from 2/18/10 at: http://www.cfids.org/webinar/series2010.asp

    I have not always been a fan of the CFIDS Assoc. but Dr. Vernon has been a significant factor in turning me into a supporter.

  • Guest

    The Dutch proposal that WPI study is limited to the Tahoe outbreak is baseless. WPI's samples were taken from over a half-dozen practitioners' inventories around the U.S., to which many patients traveled from afar. See Mikovits' presentation video on ProHealth. (What's interesting, though, is that this travel-factor that added more states and even foreign countries, Ireland for one, appears to have been initially overlooked.) You'd have to assume all those patients either visited Tahoe, or whatever happened there is far more communicable.

    Even if Klimas and Peterson helped write the Canadian ME/CFS Consensus criteria, it's the best one to rule in by neuroimmune findings and rule out other psychiatric disorders to select patients. WPI can share their primers with everyone. As long as others are using B.S. definitions, this will be tit-for-tat. Ideally, yes, we should be sharing samples. I wouldn't share any with Wessley's U.K. group without a 24-hour watch by U.S. troops; the psychosomatic folks have too much at stake to do good science, which is becoming a rarity all-around anyway.

  • Guest

    After this, you don't know what to believe. The second UK study also failed to use the Canadian Criteria; it doesn't look then that Jonathan Kerr has the same interests as his WPI grant collaborators, or that his voice didn't carry much weight on that team. That's alarming.

    Funny that Dr. Vernon would call out WPI for not sharing. The study she designed with Dr. Reeves went to Duke U., which didn't share theirs with anyone either. BTW, no one in Duke Infectious Disease will see anyone diagnosed with “CFS,” much like Emory. As far as U.S. practitioners are concerned, Fukada-CFS is a wastebasket Dx, grounds for turning away some very sick people.

    Sadly, contrary to Dr. Vernon's point, what high-fallutin' agency any of these guys hail from is irrelevant (especially in the government). There's a lot of political and monetary shuffling going on behind the scenes here and abroad. It's a sick game of chess – and we're the pawns.

  • Petra

    Wonderful! I'll be looking forward to that. Your evenhanded coverage of this topic continues to be the best on the web.

  • CBS

    Guest,

    I agree with several of your key points.

    “After this, you don't know what to believe. “

    “There's a lot of political and monetary shuffling going on behind the scenes here and abroad. It's a sick game of chess – and we're the pawns.”

    Dr. R begins the Virology 101 series by stating that it's driven by ego. Everyone wants to be the first.

    “As far as U.S. practitioners are concerned, Fukada-CFS is a wastebasket Dx, grounds for turning away some very sick people.”

    There are a very few Infectious Disease docs that appreciate a subset of CFS as a probable immune disorder characterized by frequent and numerous co-infections. They are rare.

    See the latest exchange between LA Jason and W Reeves (who on 2/14/10, was moved out of his position as Director of CFS efforts at the CDC): 'Evaluating the CDC's Empirical CFS Case Definition' – http://dps.sagepub.com/cgi/content/short/20/2/93

    Shortly after Reeves' departure (2/18/10), the CDC created an XMRV page and posted a link to that page at the top of the latest findings column on the CDC's CFS page. Kind of makes you wonder what the CDC has been finding with its division of retrovirology's work on XMRV. I understand that those results may be due out soon.

  • John

    Take questions!

  • John

    Regarding the antibody questions, is it possible that prior exposure to FeLV could generate an immune response and then upon later exposure to XMRV the body would be tricked into thinking it already had an immune response but actually XMRV would go unchecked?

    http://scienceblogs.com/erv/2010/02/when_good_a

  • dancer

    Sounds like that's what they all say. The WPI hasn't yet responded about this latest study but after the previous two UK studies, they stated publicly that neither UK group had requested materials from them.

  • dancer

    It doesn't help that the WPI publicly promotes the unpublished findings as well as the attitude that others have an agenda. (On TV, news articles, their own press releases, and FaceBook.)

  • plantecologist

    Are there legal/ethical barriers to sharing samples linked with living people?

    Great blog!

  • http://www.virology.ws profvrr

    All studies involving human subjects are strictly regulated by
    Institutional Review Boards (IRBs). Before studies can proceed, they
    must be formally written, submitted to the IRB, and approved; the
    process takes many months. Exchanging human samples would also require
    approval by the IRBs of both institutions. Exchange of samples can be
    done, but only after IRB approval.

  • Hip

    Wessely School Network

    The Nijmegen group in the Netherlands, who carried out this latest XMRV-CFS study, are part of the Wessely School network.

    Members of the Wessely School network include:

    In the US: Bill Reeves of the CDC.
    In the UK: Simon Wessely, Michael Sharpe, Anthony Cleare, Peter White, Anthony David.
    In Australia: Lloyd, Hickie, etc.
    In the Netherlands: the clinicians of the Nijmegen group.

    Mighty suspicious that the two follow up studies, in the UK and in the Netherlands, were both performed under Wessely School network influence.

    Notes on People:

    Simon Wessely often works for UNUM (a large disability insurer, and an very suspect company that has actually given a “Hungry Vulture Award” to its “deserving employees”).
    Peter White (a psychiatrist) is one of the Chief Medical Officers for Swiss Re.
    Michael Sharpe, Simon Wessely and Anthony Cleare have also worked for Swiss Re.

    Simon Wessely and Anthony David are trying to overturn the WHO formal classification of ME as a neurological disorder and to re-designate ME as a psychiatric condition.

    Simon Wessely is a Corporate Officer of PRISMA. PRISMA is being paid many millions of pounds to supply “rehabilitation” programs (such as CBT and GET) to the UK's National Health Service for use on “CFS” patients.

    Notes on Organizations:

    The majority of CFS advocacy groups are not involved in legitimate or useful advocacy. This includes: in the US: The CFIDS Association of America; in the UK: Action for ME and the ME Association; in Australia: ME/Chronic Fatigue Syndrome Society of Victoria. Most of these groups have sold out to the highest bidder. These groups benefit, either in terms of funding, influence or in other ways, by following government policy, and by their association with corporate vested interests. These groups are not helping ANYONE, except themselves.

    Sources:

    http://www.hfme.org/whobenefitsfromcfs.htm
    http://www.meactionuk.org.uk/Notes_o…ssue_in_…

  • Guest

    Prof. Racaniello, have you recorded your conversation with Prof. Goff yet? It is eagerly awaited.
    In addition, would you consider writing a post for your blog about the full suite of XMRV papers at CROI? Dr. Sharma's paper, in particular, was stellar and has generated intensive amounts of interest and discussion in many quarters.
    Thank y0u for your excellent blog.

  • http://www.virology.ws profvrr

    We were set to record last Thursday but the snow in the east foiled
    those plans. This week didn't work out for either of us, but I'm
    hopeful for next week. I won't let you down.

    As for the CROI papers – I usually prefer to discuss peer-reviewed
    articles because you never hear about the faults in presentations. But
    I'll have a look at them.

    My apologies to everyone for not being as responsive as usual lately
    in the discussions – teaching this semester is taking a big chunk out
    of my time.

  • Arabella

    I do not believe that XMRV occurs only in cluster outbreaks of CFS/ME because I have had CFS/ME for 23 years and I was not part of any cluster outbreak. I tested positive for XMRV by culture by VIP Dx in Reno, NV in December 2009. I am an example of someone who suffers from CFS/ME who tests positive for XMRV and was not part of any cluster. No one around me was suffering as I was.

  • Arabella

    I do not believe that XMRV occurs only in cluster outbreaks of CFS/ME because I have had CFS/ME for 23 years and I was not part of any cluster outbreak. I tested positive for XMRV by culture by VIP Dx in Reno, NV in December 2009. I am an example of someone who suffers from CFS/ME who tests positive for XMRV and was not part of any cluster. No one around me was suffering as I was.

  • Anonymous

    Chronic Fatigue Syndrome as reveal many criteria of disease which has found by him with some relevant idea to finding this virus……….

     hypothyroidism