Discussion in 'XMRV Research and Replication Studies' started by Jemal, May 4, 2011.
True, as far as it goes, depends on how positive a "known positive" is I suppose, she was unable to detect XMRV in a blinded sample that the WPI says is positive. That may not be the same thing as a "known positive" IF the WPI is wrong.
They didn't look for MLV's that's why they didn't find them. They used new assay's, frozen samples, They only tested two patients from the original science study. 13 negative controls and 2 positive controls were also 350 included. Unfortunately for her one of those patients was the source of a full length clone and EM showing viral budding. There are many things wrong with this paper.
I don't know... a while back Buzz had been hinting that Singh was going to be positive, then it seemed to move to hinting Singh was going to be negative. I've got no inside information here, but maybe Cort had been chatting to her?
If she is trying to find positive samples in a study, then she MUST calibrate her methods to a known positive sample. That is what they do in all other diseases. A clone will not do. Why was this not done?
It could be he has spoken with her or others that may have made certain hints...
However, the Singh study took so long I presumed it was negative as well. I had hope it would be positive though
I didn't realize he was on speaking terms with Singh. Isn't she also the one who was going to do autopsies to check for possible places in the body where the virus lives?
I don't know if he is on speaking terms with her.
And yes, she is (was?) doing the autopsy study. I am also wondering if this means the end of her research into XMRV and prostate cancer.
Surely, if she meant what she said about a possible viral cause for ME/CHF, she will still do the autopsies. If she stops now, that would look like she is not really trying to get to the answer.
I think she was doing the autopsies in relation to XMRV. If she now thinks XMRV is contamination, I don't think she will continue that research. But I am not her spokesperson, so this is just what I think and deduce
Quoting from the study:
"Using a third party phlebotomy service that collected blood samples in home visits, we obtained blinded whole blood and serum samples from 14 individuals. These individuals had repeatedly tested positive in the last two years when tested by the labs at the WPI, though this information was not available to us till the completion of our study.
The Clinical Research department at ARUP Laboratories received these specimens and processed the blood using the same protocols as for our healthy volunteers and CFS patient samples. Thus the samples were never opened in a research lab where XMRV might be present until they reached us. We tested these samples using all of the assays we developed four qPCR assays, ELISA and Western blots. None of the samples contained any evidence of XMRV."
It sounds to me like she might be implying that the WPI's lab was contaminated at some point? I don't know. All this is way above my head.
And if XMRV is contamination, how does that affect her work -- and her patents -- re XMRV and cancer?
Actually I meant this and it's from the paper. They could not even detect the 2 positives with their methods:
348 inoculated LNCaP cells with 100 ?l of plasma from 31 patients and 34 healthy volunteers, and
349 passaged the cells weekly for 6 weeks. 13 negative controls and 2 positive controls were also
350 included. Only one culture was handled at a time to prevent any cross-contamination. After
351 weeks 2, 4 and 6, cultures were lysed and analyzed by Western blots (Fig. 4) and by qPCR for
352 XMRV. No XMRV protein or DNA was detected in any of the cultures.[/B]
Yet another garbage zero-zero paper where even the author admitted that they used novel assays not able to detect XMRV. Seriously, WTF? Can we please get some true replication studies? This is becoming truly sad and pathetic; I'm beginning to lose all faith in the scientific community.
As long as scientists continue to use their own, different methods, we'll keep getting junk negative papers like this. F--king depressing.
Very good questions, especially about the patents. Maybe she thinks they are junk now? I am very interested to know if she continues XMRV research or not.
I don't think we can just go on insisting that we have known XMRV positive patients whose blood needs to be used to validate all testing.
It's looking increasingly likely that those patients thought to be positive are actually negative. This hasn't been settled yet, but it's certainly possible that the 0/0 studies are getting it right, and the other studies have had trouble with contamination.
We've yet to hear back from Silverman and Klein about the claims that some of their PC work is best explained by contamination... it seems to me that there's not much we can say about XMRV with any certainty.
Thanks Ernie. I guess I was reading a different section of the paper.
So my question is, didn't she find an XMRV - cancer connection? And if so, were different techniques used in that study or studies?
I was just reading tea leaves then. Sometimes these arbitrary patterns can fall in to place!
A riff on the Racaniello article
From Dr Racaniello - after noting all the problems with the earlier studies, Dr. Racaniello called the Singh study the 'most comprehensive study to date' and, importantly, that it took into account a number of potentially confounding factors that could have contributed to past negative results (differences in patient characterization, geographic locations, clinical samples used, and methods). That was its purpose all along. The killer here is the inclusion of 'methods' since this implies that Dr. Racaniello believes this study did all the methods right.
The clinic, of course, was Dr. Bateman's clinic - a clinician who knows how to characterize CFS patients.
They looked for XMRV using four different PCR assays and ELISA antibody and they tried to grow the virus in cell cultures; the same approaches, Dr. Racaniello reported, used by Lombardi et. al in the original paper.
As was noted many moons ago - the pol region is the most specific for XMRV since that region rarely changes in viruses. They also looked for the gag and env regions (as the WPI did). As with many of the other studies, everybody was negative via PCR and antibodies - patients and controls.
The inability to find XMRV via PCR and antibodies has, of course, happened numerous times before. This time Singh also tried to use culture - for up to six weeks - the time Dr. Mikovits stated was necessary. Unfortunately no viral protein or DNA from XMRV was found either in people in with CFS, healthy controls or people who previously tested positive for XMRV by the WPI.
Then they used the nested PCR techniques used by Lo (which the WPI endorsed). They did find some positives but concluded they came from trace amounts of mouse DNA in their reagents
Dr. Racaniello basically concluded that this study was good enough to conclude that XMRV was not found in these patients and suggested suggested that with regards to researchers the XMRV case was pretty closed but that for patients sake (non-scientists)
the WPI should look to their reagents etc. in order to clear up where the XMRV in their study came from.
Dr. Singh, Dr. Bateman and the Lights took a strong stand against the use of antiretrovirals stating that they were 'forced to conclude that prescribing antiretroviral agents to CFS patients is insufficiently justified and potentially dangerous.' and that much data suggests pathogens play a role in CFS and that work should continue.
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