Retrovirology Publishes Five Papers on XMRV and Contamination

[Cort]

Ruscetti is really the key figure here I think. He is the biggest name associated with XMRV. His lab is highly respected. Its been there been there for a long time...Its not a new fledgling Institute with a Director who can be rather outspoken shall we say..I imagine that the one researchers are waiting to hear from is Dr. Ruscetti....

 

[omerbasket]

This is all bullshit, and I wonder why some of you take it seriously. The only thing serious here is that when you say crap too much times, sometimes too much people start to believe you.

The WPI have isolated infectious XMRV from more than 600 XMRV positive PCR samples, and they have done it from both plasma and PBMCs; They sequenced every PCR product and chekced it with at least one other method to confirm to positivity of the sample; And, perhaps most importantly, they showed an immune response. I'm sorry, but people's immune system cannot respond to a lab contamination...The contamination thing is crap, crap, crap, but if people here would think of it as a real possibility, than the truth might remain sealed and people with ME/CFS and other XMRV-related diseases would continue to be sick for many more decades.

 

[August59]

That's just another piece of the puzzle isn't it? How do you explain her and her daughter's improvement and Andrea Whittemore's improvement (although we don't know of her treatment regime)...if XMRV doesn't work out? Is there another retrovirus or is it something else?

When Dr. Mikovits was doing her study and proved that PWC had approx. 30 viruses (or antibodies) as opposed to the 8-10 that the healthy controls had is proof that something is allowing these viruses and infections to proliferate or the complete malfunction of our immune system. Not that this is news, but it is very messed up if it allows all of the things indicated in the study to be happening. Again it could be a combination of these thats the problem

 

[CBS]

Ruscetti is really the key figure here I think. He is the biggest name associated with XMRV. His lab is highly respected. Its been there been there for a long time...Its not a new fledgling Institute with a Director who can be rather outspoken shall we say..I imagine that the one researchers are waiting to hear from is Dr. Ruscetti....

Frank (http://ccr.cancer.gov/staff/staff.asp?profileid=5488) or Sandra (http://ccr.cancer.gov/staff/staff.asp?profileid=5518)?

He's been a bit more visible (at least in public meetings) and he's been doing work on the ramifications or retroviral infections (eg. cytokine upregulation, etc.) but her position is also directly oriented to the question of retroviruses (Laboratory of Cancer Prevention - Head, Retroviral Pathogenesis Section).

 

[free at last]

I don't want to sound too depressing, but if XMRV is proven a contaminant, I think it will do more damage to our cause than good. You would have two retrovirus incidents in the history of this disease, the second time with very reputable scientists involved. I am not so sure many scientists would still want to get involved with us. ME/CFS might be seen as too difficult and unfortunately it's in our nature to take the paths of least resistance. The third time some scientist claims he has found the cause of ME/CFS, he will come under intense scrutiny and his reputation will be in jeopardy. It would probably be easier to just study the mating rituals of hummingbirds.

Still believing in XMRV though and I hope we'll see a response soon from the scientists who think XMRV is for real.



Ignoring it won't make it dissappear. More and more news outlets are picking up this story. British, Spanish and now German news outlets have picked up the story. I am sure US news outlets will soon follow.


http://www.guardian.co.uk/society/2010/dec/20/chronic-fatigue-me-research


http://www.aerzteblatt.de/nachricht...rsache-des-Chronischen-M_digkeitssyndroms.htm

Powerful statements like these tend to stick in people's minds.

Other people on the "other side" of the XMRV debate have also quickly picked up the news and are gloating. Our good friend ERV for example.

I agree the damage can not be underestimated Jemal

 

[CBS]

This is all bullshit, and I wonder why some of you take it seriously. The only thing serious here is that when you say crap too much times, sometimes too much people start to believe you.

The WPI have isolated infectious XMRV from more than 600 XMRV positive PCR samples, and they have done it from both plasma and PBMCs; They sequenced every PCR product and chekced it with at least one other method to confirm to positivity of the sample; And, perhaps most importantly, they showed an immune response. I'm sorry, but people's immune system cannot respond to a lab contamination...The contamination thing is crap, crap, crap, but if people here would think of it as a real possibility, than the truth might remain sealed and people with ME/CFS and other XMRV-related diseases would continue to be sick for many more decades.

I could not agree more about the number of other questions that the contamination claim fails to address (for beginners - immune responses, pictures of budding viruses - Singh - and IHC). This is far from settled. And yes, this may all be BS. However, Alter, Mikovitz, Silverman, the Ruscetti's, and Singh, amongst others, are the ones who need to put this to rest as my doctors aren't going to do anything to treat my ME/CFS as a retroviral infection based on my word (or yours or anyone else on this or any other forum) alone.

 

[markmc20001]

Hi, all.

I think it's important to note that these papers are concerned only with PCR testing. DNA contamination can impact PCR testing, but it cannot explain either the ability to culture viruses, which means that there were whole virions in the samples, or to detect antibodies, showing that the immune system had responded to a viral infection in the host.

Judy Mikovits has emphasized many times that using only the PCR technique is not sufficient for conclusively identifying the presence of these retroviruses. It's a powerful method, but it has its limitations, and sensitivity to contamination is one of them.

Best regards,

Rich

It's good to see a real scientist comment on this shocking news. Thank you.

 

[dannybex]

They sequenced every PCR product and chekced it with at least one other method to confirm to positivity of the sample; And, perhaps most importantly, they showed an immune response. I'm sorry, but people's immune system cannot respond to a lab contamination...The contamination thing is crap, crap, crap, but if people here would think of it as a real possibility, than the truth might remain sealed and people with ME/CFS and other XMRV-related diseases would continue to be sick for many more decades.

Hi Omerbasket,

This XMRV science has always been way over my head. Can you explain how or why people cannot have an immune response to a lab contaminant?

Thanks,

Dan

 

[CBS]

Unfortunately, i think this is not known. Should have been published already. Like so much, it takes longer and longer. But i think she will want to make sure she has not been working for a year or more on something that's not really a human virus. So one more group that will be working to find out what's the truth. I don't think she will just say "Oh, yes, i've been wasting my time and your money for the last year or two. Sorry, i'm not very smart, you know...".

A large majority of Dr. Singh's work has been on tissue samples and using Immunohistochemistry (IHC) to detect the actual virus itself on a slide (her picture of a budding retrovirus is the one that is often seen accompanying articles on XMRV). As has been stated elsewhere, she expected to submit her autopsy study in November. As she stated in September, reviewers have been very cautious. I don't expect that these four articles on PCR will speed anything up.

Does anyone know for sure whether anything published today has any relevance to the accuracy of IHC (potential for a false positive stain)?

 

[free at last]

This is all bullshit, and I wonder why some of you take it seriously. The only thing serious here is that when you say crap too much times, sometimes too much people start to believe you.

The WPI have isolated infectious XMRV from more than 600 XMRV positive PCR samples, and they have done it from both plasma and PBMCs; They sequenced every PCR product and chekced it with at least one other method to confirm to positivity of the sample; And, perhaps most importantly, they showed an immune response. I'm sorry, but people's immune system cannot respond to a lab contamination...The contamination thing is crap, crap, crap, but if people here would think of it as a real possibility, than the truth might remain sealed and people with ME/CFS and other XMRV-related diseases would continue to be sick for many more decades.

I want to think like you omerbasket, its just my knowledge is way to short to know the truth, i do belive this is a campaign that has been silently worked on for months in the UK, and i belive they waited to see what turned up at the BWG before releasing these papers all in a row to cause the most damaging effect, and guess what THEY HAVE wishfully thinking it away will not work.We need hard evidence and statements from the other big names, i hope the statements will be coming soon, and the evidence eventually,But those who want us to still be attending the psychiatrists, have struck a very clever blow.The news storys going around the world are proof of that. Ill say this, yes im worried because i dont understand the science, have i given up faith in Judy, NOPE not untill she says all is lost. Ill continue to trust her untill she says theres a problem, some may call it blind faith, i call it knowing which side cares about us, and which side pretends they do. I still support the WPI i support there science untill proven otherwise.

But im worried, and im down. I said before in the UK we have nothing, well worse than that, we have groups that are so intent on finding us all nuts, that the research will look for holes, rather than connecting points, and they may have found some holes that as of yet i dont fully understand the implications of, and neither will a lot of other people, its a win win situation for them i think. Only Dr sing and Lipkin can even begin to undo the damage these clever negative researchers have done. just like a london bus nothing for ages then 5 come along all at once, a coincedence i think not, its a stategy.Its about time the scientists on our side started playing these stategic games. I hope they do, if ever they get the chance. What a mess

 

[Cort]

The Huber Paper -similar outcome but different initial results; high rate of positives in healthy controls but not people with CFS - definitely at odds with prior patterns of XMRV prevalence.

Contamination of human DNA samples with mouse DNA can lead to false detection of XMRV-like sequences - Brendan Oakes…Brigitte Huber

Dr. Huber reported that her laboratory has been involved in CFS research since 2005 and thus has a substantial laboratory of samples. First Dr. Huber used quantitative PCR assay (WPI used nested PCR assay) that was specific for XMRV (ie did not cross react with any sequence known to occur in mouse DNA) provided to her by Dr. Singh. This assay, however, did not pick up any XMRV in the hundred 48 samples tested. Questioning whether she needed to search for more divergent MLV sequences she then turned to the nested PCR used by the WPI in their study.

Now she found that almost half healthy controls 19/36) and but only a small # of people with CFS (2/112) tested positive. When they started testing the samples (n=158) they found that all the samples that tested positive for XMRV also tested positive for mouse DNA. Most of the samples, on the other hand, that tested negative for XMRV from/MLV’s were negative for mouse DNA. This suggested to her that they were actually picking up mouse DNA rather than a human virus.

When she sequenced this group she found a very high degree of diversity - indicating 15 different virus strains have been uncovered; three healthy controls appeared to have XMRV but the rest had sequences identical to or closely related to endogenous MLV's (ie mouse contaminants ). Her genetic sequence indicated three clusters for present, each of which contained endogenous sequences of a different subtype.

Contamination? She then ran two tests for contamination; a qPCR assay and the IAP test mentioned in the other paper and found that many of the CFS and control samples were positive for mouse sequences using these tests. (The high percentage of CFS patients positive for mouse DNA could have derived from when those samples were processed…On the other hand….is there a reason people with CFS might have higher numbers of mouse DNA?) Almost all the samples that had putatively been tested positive for XMRV using the WPI’s primer tested positive for contamination using either test (IAP - 100% accurate).

Conclusions - She reported that her data suggests that the mouse DNA contamination most likely occurred early in the process during blood collection, isolation of the PBMC’s, or the preparation of the DNA. She points out several times that her negative controls were always negative despite many rounds of testing - indicating that the contaminants did not come from her laboratory. She has not been able to pinpoint a specific reagent or laboratory vessel that is positive for mouse DNA. Preliminary experiments suggest that a serum and a saline solution could have been responsible - of which varying amounts of mouse DNA have been present in different batches. .

Huber/Alter Sequences Similar - Rather disturbingly Dr. Huber reported that the sequences that showed up at her laboratory and those found in the Alter study were ‘similar’ and were closely related to endogenous retroviruses. (A retro virologist I spoke to noted a similar caution).

 

[jeffrez]

posted in other thread.

 

[Cort]

A large majority of Dr. Singh's work has been on tissue samples and using Immunohistochemistry (IHC) to detect the actual virus itself on a slide (her picture of a budding retrovirus is the one that is often seen accompanying articles on XMRV). As has been stated elsewhere, she expected to submit her autopsy study in November. As she stated in September, reviewers have been very cautious. I don't expect that these four articles on PCR will speed anything up.

Does anyone know for sure whether anything published today has any relevance to the accuracy of IHC (potential for a false positive stain)?

I have no idea but Huber's study found that Singh's XMRV assay did not pick up any XMRV. All the XMRV positive results she got using the WPI assay also tested positive for contamination using the IAP test. On the other hand there is a big discrepancy that is hard to figure out; it was the healthy controls that tested positive - not the CFS patients. Since the samples were collected at different times this could have been from something that was inadvertently introduced during the sample collection for the controls....

I think the virus is there? I think Hue is saying that it originated from an endogenous retrovirus in a mouse cell line........this is the really confusing part.......is it bits of mouse DNA that look like XMRV or is it XMRV? XMRV is, after present.......they can and do grow it out of the 22RV1 cell line...That's how they are testing these various drugs, etc.

.Hue thinks it was passed into that cell line by being passaged through mice (whatever that means)....THen, I suppose, the cell line itself is infecting laboratories? Does it get into a prostate cancer cell from a lab or does the person actually get XMRV from some other way? Its confusing, that's for sure.

 

[currer]

Hi all.
I just want to point out that the release of these three papers the week before Christmas has been cleverly calculated to give them maximim publicity impact as no work can be done to refute their ideas until after the holiday - this means a good three weeks of impact.
The politics of CFS/ME never fails to intrigue me!
It is surprising how eagerly this has been reported in the UK when the Alter paper was published without a peep from the media.

For what it's worth I do not feel that these new papers will have the last word.
We must and can fight on. As Alter said our goal is to find the truth - and that will show we are genuinely ill.

 

[hope love light]

New Study Dismisses Link of XMRV to CFS!!

Thought you might like to read this!!

LONDON A virus previously thought to be linked to a baffling condition known as chronic fatigue syndrome is not the cause of the disease, scientists said on Monday after their study found previous research was contaminated in the lab.

Researchers from University College London, the Wellcome Trust Sanger Institute and Oxford University said cell samples from patients in earlier studies were contaminated with the virus, known as XMRV, which is found in the DNA of mice.

http://www.msnbc.msn.com/id/40751132/ns/health-health_care/

 

[Jemal]

Yeah, we have a 10 page discussion about it here:
http://forums.aboutmecfs.org/showth...blishes-Five-Papers-on-XMRV-and-Contamination

 

[Jemal]

I could not agree more about the number of other questions that the contamination claim fails to address (for beginners - immune responses, pictures of budding viruses - Singh - and IHC). This is far from settled. And yes, this may all be BS. However, Alter, Mikovitz, Silverman, the Ruscetti's, and Singh, amongst others, are the ones who need to put this to rest as my doctors aren't going to do anything to treat my ME/CFS as a retroviral infection based on my word (or yours or anyone else on this or any other forum) alone.

This! We need them to free us.

 

[Cort]

And, perhaps most importantly, they showed an immune response. I'm sorry, but people's immune system cannot respond to a lab contamination...The contamination thing is crap, crap, crap, but if people here would think of it as a real possibility, than the truth might remain sealed and people with ME/CFS and other XMRV-related diseases would continue to be sick for many more decades.

That is the antibody test - which has always been Dr. Mikovits biggest backup....So it would seem that their job now, as it has always been, really, is to show that the antibody is completely specific for XMRV; that its not cross-reacting with anything else. Dr. Mikovits and Dr. Bagni have reported progress in this area. Dr. Singh has her own antibody test....This was all about PCR.

 

[jeffrez]

Maybe it's because XMRV doesn't in fact cause CFS. Nothing so far has shown that it does, at least. I know many people want to believe it does, but believing in something doesn't make it true. At this point the most we can say is maybe it does, and maybe it doesn't. The best the strongest proponents can even show is that it's possibly associated. Personally I doubt any direct or sole causation will be found - it just doesn't seem likely given the complexity of CFS. And it might still play some minor or additive role, so that treating it where it exists improves symptoms. But the weight of the evidence so far suggests that it doesn't cause the disorder.

Not sure why people would be upset with research just because it doesn't show what they want it to, other than the initial disappointment that will come when we find out XMRV doesn't cause CFS, which is understandable. We're best served in the long run by finding out what really causes CFS, not by clinging to false hopes despite the evidence to the contrary.

 

[August59]

Did HIV face this much resistance??

Trial Management Group (TMG)

The Trial Management Group (TMG) will be responsible for the day-to-day running and management of the trial. It is composed of:

• The three principal investigators

1. Professor PD White, Professor of Psychological Medicine, Centre for Psychiatry, Wolfson Institute of Preventive Medicine, Bart's and the London, Queen Mary School of Medicine and Dentistry, Department of Psychological Medicine, St Bartholomew's Hospital, London, EC1A 7BE.

2. Professor MC Sharpe, Professor of Psychological Medicine and Symptoms Research, School of Molecular and Clinical Medicine, Symptoms Research Group, Royal Edinburgh Hospital, Edinburgh, EH10 5HF.

3. Professor T Chalder, Professor of Cognitive Behavioural Psychotherapy Academic Department of Psychological Medicine, Guy's, King's and St Thomas' School of Medicine, Weston Education Centre, Cutcombe Road, London SE5 9RJ

• All centre leaders and co-leaders
1. Dr D Wilks
2. Professor S Wessely
3. Dr M Murphy
4. Dr BJ Angus
5. Professor T Peto
6. Dr E Feldman
7. Dr G Murphy
8. Hazel O'Dowd
------------------------------------------------------------------------------------

This study is a crock anyway from the point of it's source. These are all psychology "grunts" that are doing nothing more than trying to butter their bread.

Did HIV face this much doubt and resistance? Something feels very wrong with this "UK" group, plus "UK" based journal spreading junk. These papers are so inflated with BS that you can't follow them.

Could it be that the UK has something to hide? I'm not much on conspiracy theories, but something still bothers me about the sealed documents they have! It's not so much that they are sealed, but why the extrordinary length of time.

This whole damn contaimination thing is way beyond what it should conceivably be and what it should have ever been. McClure (who by the way looks like the wicked witch from Wizard of Oz) and Stoye (why is that SOB even in the US, revoke his damn visa) are just extraordinarly focused on this contamination view).

The real professionals can't even get their, badly needed for sake of patients, studies published in journal. How is it that these psychologist can get 5 studies published at one time. When and if the truth ever comes out about this I hope they can be prosecuted for "Crimes against Humanity" and pump them full of drugs and let them spend the rest of their years in a pshyco ward!!!!