XMRV CFS UK study #II

[Gerwyn]

Gerwyn,

judym @wpinstitute.org
(I put a space before the @)

thank you Julian much appreciated Can I send my summary post to this address and if so how do I attach it. In this area I am hopeless!

 

[Sasha]

thank you Julian much appreciated Can I send my summary post to this address and if so how do I attach it. In this area I am hopeless!

Hi Gerwyn - why don't you just copy and paste your summary into the body text of an email to Dr Mikovits with a short explanatory note at the top?

ETA: If you just want to tell her that additional point about the controls, wouldn't it be better just to send her a brief email about that, given that you think she's covered the other issues in your summary? We don't want her spending time reading emails that she could be spending in the lab!

 

[julius]

thank you Julian much appreciated Can I send my summary post to this address and if so how do I attach it. In this area I am hopeless!

You could just copy and paste it into your email. If you don't know how, let me know I can tell you.

 

[anne]

When the WPI study was published there was considerable debate on this forum regarding our hopes and fears for future replications studies. Posters were anxious that anyone undertaking research into XMRV and ME should be aware of the 'political' backgroud to ME, specifically on the issue of the cohorts selected for study. Many of us believed that only a prospective study of Canadian defined subjects was likely to replicate WPI's findings. Dr Bell's involvement (albeit possibly in a minor role) in the current study is to be welcomed in this respect.

Marco, Dr Kerr, not Dr. Bell, oui?

 

[garcia]

I have been thinking about the question of cohort selection and have found a way to rule it out as a possible confounding factor.

We know the VIP tests are about 50% positive in the UK. And what is the 'cohort' of people getting that test? A bunch of people who think they probably have CFS. Not well defined at all. In fact, you really can't get a more loosely defined 'cohort'.

So, how could the UK researchers pick a cohort with ZERO positives?

1) Random Chance.
But the probability of this is so infinitesimally small, it is just not worth considering.

All of the above suggests that the cohort-issue, whilst important in general terms, is actually a bit of a red-herring here, and takes us away from the key-issue which is methodology. Method, method, method.

 

[julius]

Yep. Let's check cohort off the list. Definitely method.

 

[Abraxas]

All of the above suggests that the cohort-issue, whilst important in general terms, is actually a bit of a red-herring here, and takes us away from the key-issue which is methodology. Method, method, method.

I agree Garcia.

 

[Marco]

Marco, Dr Kerr, not Dr. Bell, oui?

Oui. Merci et desole a tous!

 

[ukxmrv]

So that leaves (?)

1. Collection method unknown
2. Frozen patient samples (stored for years)
3. Lack of LNCaP co-culture for 5 days prior to PCR

what are 4, 5, 6 etc?

 

[Cort]

Its always been method. You can't get zero results from large patient studies by choosing the wrong cohort- unless the WPI is super selective in their patient selection process - and they've stated that they haven't been. I think this is the key

Careful reading of the Science paper shows that increasing the amount of the virus by growing the white blood cells is usually required rather than using white blood cells directly purified from the body.

Although honestly how the Retrovirology missed this I don't know. If they knew about it and ignored it they must have assumed that their techniques would have been able to pick up the vanishingly small levels of XMRV in unstimulated cells. That study cost alot of money!

You need to stimulate the cells first but check this out - they said that growing the white blood cells is usually required - not always. Since the UK study used what Dr. Vernon suggested were more sensitive techniques then you would have thought they would have picked up the virus at some point - since according to this statement it isn't always necessary to grow the white blood cells - but they obviously didn't. Perhaps the other factor the WPI listed came into play.

One the problems is that this appears to be an unusual bug - you can't do 'normal' work and find it. Somebody, either the DHHS group or the CDC or somebody else is going to follow the WPI's work to the letter and then we'll know.

 

[Elliot]

I love you guys :')
My heart literally sank when I first read the news, as this xmrv has been keeping me going! It's great to have people with a biological background to explain and analyse everything.

 

[Knackered]

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

 

[anne]

One the problems is that this appears to be an unusual bug - you can't do 'normal' work and find it. Somebody, either the DHHS group or the CDC or somebody else is going to follow the WPI's work to the letter and then we'll know.

Which raises your questions again, Cort. Why the heck didn't they? I'm not going to believe in this group that it's bad intention. Why assume you can find it using other methods? And why then, proclaim, as the first UK study did, "If it was there, we would have found it?"

 

[Quilp]

'Perhaps the most important issue to focus on is the low level of XMRV in the blood. XMRV is present in such a small percentage of white blood cells that it is highly unlikely that either UK study’s PCR method could detect it using the methods described'

As a non scientists this is the gem that I was looking for. Basically almost no chance of finding the virus using these methods.

Mark

 

[Frickly]

I and apparently many others cannot access the WPI website. Could someone copy and paste the the news in the WPI link above .

Thanks,

 

[Countrygirl]

=Frickly;47667]I and apparently many others cannot access the WPI website. Could someone copy and paste the the news in the WPI link above .

Thanks.

Me too. I haven't been able to access it for 24 hours.

 

[Gerwyn]

Its always been method. You can't get zero results from large patient studies by choosing the wrong cohort- unless the WPI is super selective in their patient selection process - and they've stated that they haven't been. I think this is the key



Although honestly how the Retrovirology missed this I don't know. If they knew about it and ignored it they must have assumed that their techniques would have been able to pick up the vanishingly small levels of XMRV in unstimulated cells. That study cost alot of money!

You need to stimulate the cells first but check this out - they said that growing the white blood cells is usually required - not always. Since the UK study used what Dr. Vernon suggested were more sensitive techniques then you would have thought they would have picked up the virus at some point - since according to this statement it isn't always necessary to grow the white blood cells - but they obviously didn't. Perhaps the other factor the WPI listed came into play.

One the problems is that this appears to be an unusual bug - you can't do 'normal' work and find it. Somebody, either the DHHS group or the CDC or somebody else is going to follow the WPI's work to the letter and then we'll know.

Hi cort .Our baby is more like a latent virus if blood was a party aids would be dancing on the tables in full view xmrv would be hiding in the kitchen in a cupboard and would only come out if pulled then pushed or activated in some other way

 

[rebecca1995]

I feel a lot better after reading this! I think it's a great response.

Onward and upward, folks! :victory:

 

[rebecca1995]

WPI Response to Second UK Study

https://www.wpinstitute.org/news/news_current.html


February 18, 2010: WPI is aware of the recent UK study that was unable to detect the presence of XMRV in any CFS patient samples. Although researchers at the WPI were not involved in this project, our work in XMRV continues with researchers around the world. We look forward to the results of studies which replicate the methods used in the original research described in the journal Science in October, 2009.

Information Regarding XMRV Studies

1. The authors of the Science paper established the existence of XMRV as an infectious human blood borne retrovirus for the first time in blood of patients diagnosed with Chronic Fatigue Syndrome (CFS). Previous studies had established the presence of XMRV sequences and protein in human prostate tissue.

2. In the Science paper, the presence of XMRV in well-characterized patients with CFS was established using multiple technologies:

a) PCR on nucleic acids from un-stimulated and stimulated white blood cells;
b) XMRV protein expression from stimulated white blood cells;
c) Virus isolation on the LNCaP cell line; and
d) A specific antibody response to XMRV.

3. The authors of the two UK studies did not attempt to replicate the WPI study. Replication requires that the same technologies be employed. The WPI sent reagents and information to several groups of researchers in an effort to support their replication studies. Neither UK study requested positive control blood, plasma or nucleic acids from the WPI.

4. The collection, preparation and storage of DNA were completely different between the Science and UK papers. The latter studies do not show data on blood harvesting or storage. Nor do the studies disclose the quantity of isolated cells. Insufficient number of cells analyzed may result in failure to detect a low copy virus like XMRV, regardless of the sensitivity of the assay. Neither UK study provides detail to allow interpretation of how many white blood cells were analyzed.

5. Patient population selection may differ between studies.

6. The UK authors were unable to detect XMRV, even though 4% of healthy individuals were found to be infected in the US. Japanese scientists detected XMRV in 1.7% in healthy blood donors in Japan. The two previously identified human retroviruses have distinct geographical distributions.

7. Perhaps the most important issue to focus on is the low level of XMRV in the blood. XMRV is present in such a small percentage of white blood cells that it is highly unlikely that either UK studys PCR method could detect it using the methods described. Careful reading of the Science paper shows that increasing the amount of the virus by growing the white blood cells is usually required rather than using white blood cells directly purified from the body. When using PCR alone, the Science authors found that four samples needed to be taken at different times from the same patient in order for XMRV to be detected by PCR in freshly isolated white blood cells. More importantly, detection methods other than PCR showed that patients whose blood lacks sufficient amount of XMRV detectable by PCR are actually infected. This was proven by the isolation of viral proteins and the finding of infectious XMRV isolated from the indicator cell line LNCaP. The authors of the Retrovirology paper admit that their neutralization assay did not detect bacterially expressed XMRV gag and that positive control sera was needed to validate their assay. The WPIs monoclonal antibodies specifically and sensitively completed the immune response demonstrating the assays sensitivity and specificity for XMRV envelope.

Simply stated the only validated reliable methods for detecting XMRV in CFS patients, to date, are the methods described in Science. Failure to use these methods and validated reagents has resulted in the failure to detect XMRV. A failure to detect XMRV is not the same as absence of this virus in patients with CFS.

 

[Knackered]

Edit......