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German Study - No XMRV in CFS & MS

Discussion in 'XMRV Research and Replication Studies' started by shannah, Dec 27, 2010.

  1. shannah

    shannah Senior Member

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    Source: PlosOne
    Vol 5, #12 (Preprint)
    Date: 24 december 2010
    URL:
    http://www.plosone.org/article/fetc...leURI=info:doi%2=10.1371/journal.pone.0015632

    Edit:
    try instead

    http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015632


    No evidence for XMRV in German CFS and MS patients with fatigue
    despite the ability of the virus to infect human blood cells in
    vitro
    ---------------------------------------------------------------
    Oliver Hohn(1,5), Kristin Strohschein(2), Alexander U. Brandt(3),
    Sandra Seeher(1), Sandra Klein(1), Reinhard Kurth(4), Friedemann
    Paul(3), Christian Meisel(2), Carmen Scheibenbogen(2,#), Norbert
    Bannert(1,5,#,*)
    1 Centre for Biological Security 4, Robert Koch-Institute, Berlin,
    Germany,
    2 Institute for Medical Immunology, Charite - Universitatsmedizin
    Berlin, Berlin, Germany,
    3 NeuroCure Clinical Research Center (NCRC), Charite Universitats-
    medizin Berlin, Berlin, Germany,
    4 Robert Koch-Institute, Berlin, Germany,
    5 Centre for Retrovirology, Robert Koch-Institute, Berlin, Germany
    * E-mail: bannertn@rki.de
    # These authors contributed equally to this work.


    Received: September 16, 2010; Accepted: November 18, 2010; Published:
    December 22, 2010


    Abstract

    Background
    Xenotropic murine leukemia virus-related virus (XMRV), a novel
    human retrovirus originally identified in prostate cancer
    tissues, has recently been associated with chronic fatigue
    syndrome (CFS), a disabling disease of unknown etiology
    affecting millions of people worldwide. However, several
    subsequent studies failed to detect the virus in patients
    suffering from these illnesses or in healthy subjects. Here we
    report the results of efforts to detect antibody responses and
    viral sequences in samples from a cohort of German CFS and
    relapsing remitting multiple sclerosis (MS) patients with
    fatigue symptoms.

    Methodology
    Blood samples were taken from a cohort of 39 patients
    fulfilling the Fukuda/CDC criteria (CFS), from 112 patients
    with an established MS diagnosis and from 40 healthy donors.
    Fatigue severity in MS patients was assessed using the Fatigue
    Severity Scale (FSS). Validated Gag- and Env-ELISA assays were
    used to screen sera for XMRV antibodies. PHA-activated PBMC
    were cultured for seven days in the presence of IL-2 and DNA
    isolated from these cultures as well as from co-cultures of
    PBMC and highly permissive LNCaP cells was analyzed by nested
    PCR for the presence of the XMRV gag gene. In addition, PBMC
    cultures were exposed to 22Rv1-derived XMRV to assess
    infectivity and virus production.

    Conclusion
    None of the screened sera from CFS and MS patients or healthy
    blood donors tested positive for XMRV specific antibodies and
    all PBMC (and PBMC plus LNCaP) cultures remained negative for
    XMRV sequences by nested PCR. These results argue against an
    association between XMRV infection and CFS and MS in Germany.
    However, we could confirm that PBMC cultures from healthy
    donors and from CFS patients can be experimentally infected by
    XMRV, resulting in the release of low levels of transmittable
    virus.

    --------
    (c) 2010 PlosOne
     
  2. shannah

    shannah Senior Member

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    This study came through Co-Cure this morning. I've copied it exactly as reported but the link doesn't appear to be working.
     
  3. shannah

    shannah Senior Member

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    Can one of our scientific minds critique this study for us please.
     
  4. alex3619

    alex3619 Senior Member

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    Logan, Queensland, Australia
    Hi everyone, I haven't read the entire paper yet, but I dismissed this paper as probably not important when I saw it on cocure. First, it was published in PLoS: this is a low rating journal. Second it uses nested PCR, which we already know is problematic. Third, while they did culture for the virus they only cultured for 7 days: we know the WPI finds it takes up to 42 days to culture the virus. On the other hand, they did use Fukuda at least for the CFS diagnoses.

    If I get around to reading this paper, which might not be soon, I will post a longer commentary.

    Bye,
    Alex

    ps Almost forget, isn't this another zero zero study? They did however show that XMRV can infect these cultures and is an infective virus. This may be worth a more detailed look.
     
  5. shannah

    shannah Senior Member

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    Thanks Alex for taking a look and commenting as quickly as you did.

    I knew enough not to be alarmed by it but certainly not enough to understand why not.

    In the Nevada Newsmakers clip just before Christmas, I noticed Judy is now saying it can take up to 45 days to culture, up from her previously stated 42.
     
  6. alex3619

    alex3619 Senior Member

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    Logan, Queensland, Australia
    Hi shannah, thanks for this, somehow I missed noticing that detail from Judy - my brain probably heard 42. I wonder if this number is going to keep going up? If extra culturing time increases the capacity to find the virus enough then it may become standard for XMRV testing. Bye, Alex
     
  7. jeffrez

    jeffrez Senior Member

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    NY
    So how come they didn't have antibodies?
     
  8. kurt

    kurt Senior Member

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    Syracuse, Utah, USA
    The original Science article study stated the T and B cells were cultured for 42 days. She can't change that to 45 days now, other labs should be able to find what she found the same way she found it. And even then, they now must run the more robust contamination studies, which WPI should also be doing as well, given the results of the Retrovirology studies. If WPI does not re-run their samples with the more robust contamination studies, they will lose credibility in the scientific community.
     
  9. alex3619

    alex3619 Senior Member

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    Logan, Queensland, Australia
    Hi kurt, I agree, this is a real risk. I think that it will happen (the new tests that is), but it will take a while. I also worry that they need to test their culture medium and other reagents with the alternative mouse DNA tests. Bye, Alex
     
  10. alex3619

    alex3619 Senior Member

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    Logan, Queensland, Australia
    Hi Mr. Kite, there might be a number of reasons. They didn't use XMRV positive human controls to validate their methods. Their antibody methods were from a prior zero zero study, and validated using controls in yet another zero zero study. The goat serum controls were probably from recently infected goats. Why does everyone forget/ignore that the virus disappears from the blood after a number of days? Their antibody tests were to directly find XMRV using XMRV targeted manufactured antibodies, they did not look for natural antibodies to XMRV.

    They found when they infected a culture that XMRV was detectable using their PCR test after seven days. Their test does work on tissue that has a heavy viral load.

    The good news: reverse transcriptase. They found high RT in the infected culture, but within expected range for controls and CFS patients. In other words the virus is not massively replicating in CFS patients. This is a good thing, but we already kind of knew this from the low copy number issue in CFS patients.

    Take home points: their tests should have been expected to not work based on prior studies. They did however show, again, that this is a real virus and not strongly present in the blood, nor massively replicating.

    Bye, Alex
     
  11. Cort

    Cort Phoenix Rising Founder

    I've got to look at the paper too but I do believe that nested PCR is what the WPI used in their original study.

    I think this is an old study though.....
     
  12. shannah

    shannah Senior Member

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    Hi Cort,

    This particular study was
    Received: September 16, 2010; Accepted: November 18, 2010; Published: December 22, 2010
     
  13. jeffrez

    jeffrez Senior Member

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    Thanks for the explanations. I'm having a hard time believing antibodies wouldn't be detected if they existed in the samples, but I suppose it's possible their methods were flawed. There always seems to be an "out" to keep the yea-sayers going (and the money flowing in to WPI, et al. ;-)), but hopefully it will be sorted THIS year. Although perhaps that's as overly optimistic as expecting it to have been sorted this past year. Once that gravy train gets rolling, I'm sure it must become very compelling for these researchers to maintain their "rivalry" for as long as possible.
     
  14. omerbasket

    omerbasket Senior Member

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    Kurt, I know you are against XMRV and the WPI - but when Judy talked about 45 days of culture in norway, she did so when she presented it to patients, and she probably just rounded the number up. Anyway, what they really are saying is that you need at least 21 days of culture, and at most 42 days.
    WPI did almost everything that can be done in order to check for contamination. All of those papers about contamination talked about PCR. But the WPI didn't use only PCR in their study. They used a number of methods, and they showed an immune response to the virus - and all of those papers have chosen to ignonre it.
     
  15. omerbasket

    omerbasket Senior Member

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    I'm bringing here some of Gerwyn's stuff from the other forum - and you can all see for yourselves why this study, again, says nothing at all:
    About not using the technique that lead Hohn to find XMRV in respiratory tract secretions:
     
  16. eric_s

    eric_s Senior Member

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    I don't care about that kind of negative studies too much anymore. I'm waiting for BWG results or work by those people who have reported to be able to find XMRV like Alter, Lo, Hansen and hopefully Singh (the WPI too, of course, but we already know that). As long as we don't get a negative study in a journal like Science, Nature etc. it does not change much for me.
     

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