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XMRV Study No. 4

Discussion in 'XMRV Research and Replication Studies' started by Orla, Feb 25, 2010.

  1. V99

    V99 *****

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    Angela, I completely agree. I was only using very basic logic to demonstrate how use of the Oxford criteria helps no one.

    Can anyone tell me if, in the early 90's, the Ramsey ME definition could have been used? I know the Holmes definition could have been used, but what about in the UK?
     
  2. garcia

    garcia Aristocrat Extraordinaire

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    I think you are underestimating the difficulty of the science. It is rocket science, at least right now. If it was easy to detect, we would have found it a long time ago. Clearly whatever causes ME/CFS (if it is a single thing) is not easy to detect, almost by definition.

    It is true that there should be many different test designs detecting the virus, but that doesn't mean there won't be 1000 times more test designs which don't.

    Just look at the history of Helicobacter Pylori. It was discovered using new staining techniques (existing techniques wouldn't have found it). Even then it took years before there was "consensus". Not so much due to scientific reasons, but political reasons.
     
  3. Mithriel

    Mithriel Senior Member

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    These are just what I have stored on my computer. I don't know if there was a set definition, but there was definitely a concensus.

    ME was accepted by the WHO from 1969.

    This book is where he says ME has the cardinal symptoms of an abnormal response to exercise, muscle fatiguability, malaise and a prolonged recovery; muscle pain; and a variability of symptoms with time, hourly, daily, weekly. yearly that is not seen in other illnesses. He also describes all the symptoms seen before.


    (thanks to Margaret Williams)

    There were good descriptions of ME available.

    The Kerr study I think could be said to be a validation study, he used patients he had been working on, but we must ask why the other two studies were done the way they were. There was no need to rush out a study, there was no need to use such old blood using such a loose definition. They chose to do it that way.

    Can anyone really believe they sat down to decide the best way to see if they could find XMRV in CFS and decided that 19 year old blood was the way to go? If XMRV is confirmed in CFS it might be a useful exercise to check out old blood to see if it was present then, but as a starting point? When they were looking to see if HIV caused AIDS did anyone decide to use blood from homosexual men in 1960 over fresh blood from the present?

    These are people who believe (despite the evidence) that CFS is a FALSE illness belief. Patients believe they have a disease but this is false, they do not have anything wrong with the way their bodies work, no infection, nothing. It is the equivalent of a false belief that aliens are talking to you through the TV set.

    Maybe they are honourable men but they have to be prejudiced, even if unconsciously, against finding that their theories are wrong and this IS a virus caused disease. Their work over the last decades will be wrong, their status in the academic world will plummet.

    Mithriel
     
  4. natasa778

    natasa778 Senior Member

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    Why did it take so long to figure out HIV?


    I agree with Garcia on this
     
  5. Angela Kennedy

    Angela Kennedy *****

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    Hi V99 - oh yes, of course, I see what you were doing there. It is the actual speculative projections themselves elsewhere, dressed up as accurate calculations when they are not, not because of number miscalculations, but because of mistaken assumptions around what is being sampled, and the resulting inappropriate extrapolations, that are the problem, of course.
     
  6. Gerwyn

    Gerwyn Guest

    This is how careful you have to be to end up with good quality useable DNA for virology purposes
    Protocols and Procedures

    The GPCL Specimen Processing Core Laboratory uses standard operating procedures and optimized conditions. In order to see that all samples are processed in a timely fashion, and to maximize the probability of high quality DNA we suggest that the following guidelines be followed.

    Blood Collection
    For DNA extraction, collect blood in EDTA (purple-top) tubes. Vacutainer Blood Tubes from Becton Dickinson are recommended. For serum, collect blood in Serum Separator Tubes, SST (red-gray top) tubes. Vacutainer Blood Tubes from Becton Dickinson are recommended.

    Dutch study not even close did not even use the correct buffer

    Blood Tube Labels
    All sample tubes must be labeled with the following information in order to be processed: the Study Name, PI Name, Study Reference ID, and the Date Collected. Any other pertinent information regarding the sample must be noted, i.e. infectious status (if known). A computerized spreadsheet indicating this information must be enclosed with each shipment. See notification section in Scheduling/Shipping for further details.

    Tissue / Cell Collection
    Please contact the Specimen Processing Supervisor for information regarding tissue and cell collection guidelines and extraction methods.

    IRB Considerations
    Any human sample collected must be done so following all relevant IRB restrictions and HIPAA regulations.

    DNA Extraction Method
    DNA is extracted from samples according to the manufacturer's instructions using the Gentra Puregene DNA Isolation Kit. The extraction is based on a modified salt precipitation method. The dutch study did well here!!!

    Quantitation / Qualitation
    Once DNA is isolated, the DNA quantity and quality is checked using a spectrophotometer. The concentration and purity is measured by reading the Absorbance at 260 nm and 280 nm wavelengths. The expected range of high quality DNA preparations based on the A260/A280 nm wavelength ratio should be 1.7-2.0. Should the ratio deviate from this range, it may indicate protein or RNA contamination, and the sample would require repurification. do you recognise this in the dutch study?

    Sample Storage
    Following extraction the DNA is stored at 4C in DNA Hydration Solution, which is supplied with the Gentra Puregene DNA Isolation Kit. or would it be allright at -190 degrees?

    Reporting
    The Investigator is provided with a printout and/or computer copy of the Absorbance at 260 nm and 280 nm wavelengths, A260/A280 nm wavelength ratio, the DNA concentration and the total yield of DNA.

    Scheduling and Shipping

    In order to ensure timely processing of your samples and the quality of the DNA that we are able to extract, please conform to these basic shipping and scheduling guidelines. time from taking blood to processing is vital for DNA integrity

    Contact us
    Before any samples may be shipped, the study and investigator must be registered with GPCL. If you have not yet contacted us, you may contact us by phone, email or register as a user at our web site registration page.

    anyone still think that there could not be problems with DNA quality in blood taken in Holland 20 years ago?.The WPI used blood from a biobank that is a completely different kettle of fish extraction and storage was handled with great care.
     
  7. Gerwyn

    Gerwyn Guest


    absolutely bang on
     
  8. Gerwyn

    Gerwyn Guest


    absolutely bang on
     
  9. Cort

    Cort Phoenix Rising Founder

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    Thanks Gerwyn, you provide me with hope. I hope you're right :)
     
  10. hensue

    hensue Senior Member

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    thank you Garcia and Gerwyn.. Apparently it is rocket science that is such a great statement!! Whatever causes Me/Cfs is not easy to detect!
     
  11. Esther12

    Esther12 Senior Member

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  12. Marco

    Marco Old blackguard

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    Or that you can't talk or exercise a prostate gland back to being non-cancerous?;)
     
  13. Mithriel

    Mithriel Senior Member

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    These studies are working with virus not looking for it.

    Mithriel
     
  14. Gerwyn

    Gerwyn Guest

    they are culturing first andor using live tissue
     
  15. Abraxas

    Abraxas Senior Member

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    Esther, in another thread on the same study you linked to in your post (Cellular restriction factors and XMRV) subtr4ct posted this:

     
  16. ukxmrv

    ukxmrv Senior Member

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    On the subject of attempts to find the virus. I've now been sent the abstract for the Japanese blood donor study and have added it to the thread on the topic

    THE PREVALENCE OF XENOTROPIC MURINE LEUKEMIA VIRUS-RELATED VIRUS IN
    HEALTHY BLOOD DONORS IN JAPAN

    Rika A. Furuta1, Takayuki Miyazawa2, Takeki Sugiyama3, Takafumi Kimura1, Fumiya
    Hirayama1, Yoshihiko Tani1 and Hirotoshi Shibata1


    http://forums.aboutmecfs.org/showthread.php?3238-Who-found-XMRV-in-Japan/page2
     
  17. Gerwyn

    Gerwyn Guest

    Thankyou Cort This bug has defied the most sophisticated Virological detection methods since its "jump".The science that originally found it was impeccable.I am pretty confident that unaided PCR wont find it because it never has.

    As you know these days I am a baby psycho--if i ever pass my medical again.One of the best definitions of insanity i have ever heard is " to carry on doing the same thing and expecting the outcome to be different"

    I hope that the significance of the PCR negative but culture positive results will slowly seep into the mind of European retrovirologists--now that isnt rocket science
     
  18. Esther12

    Esther12 Senior Member

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    I noticed that. They also mentioned XMRV being possibly related to CFS, whereas a lot seem to only mention prostate cancer.

    @ Gerwyn: so the person who did the negative CFS study using one set of techniques was also doing this study using different techniques which worked? This is making my head ache. Why would they do this?
     
  19. Gerwyn

    Gerwyn Guest


    this is what i,m SOOO keen to find out
     
  20. natasa778

    natasa778 Senior Member

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    No not really. Scientist at present are at the very beginning of understanding about what could potentially prevent XMRV from establishing itself in host, let alone what SHOULD do that.

    Only AZT of HIV drugs so far has shown some inhibitory activity, and that was a) in vitro b) even if it inhibits xmrv in vivo in the same way /same degree that it does inhibit HIV, you are still a universe away from absolute "prevention of infection".
     

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