Article: From the PCR Side: the Cooperative Diagnostics XMRV Interview with Dr. Brent Satterfield

Article: From the PCR Side: the Cooperative Diagnostics XMRV Interview with Dr. Brent

You can view the page at http://forums.aboutmecfs.org/content.php?376-From-the-PCR-Side-the-Cooperative-Diagnostics-XMRV-CFS-ME-CFS-Interview-with-Dr.-Brent-Satterfield

 

I heard a peep out of Alter on the NIH webcast Demystifying Medicine — Chronic Fatigue Syndrome: Is There a Virus? held February 22, 2011.

Both Alter and Lo were inspiring. (Gill was something else.)

You can watch it and look at the slides here.

Thanks for that, gracenote... I forgot about that video... I was going to watch it but then I couldn't face sitting through 2 hours of video!
But if it's worth watching, then I might try to.

 

I was going to watch it but I couldn't face sitting through 2 hours of video!

yes, that plus something called a Fair Access Policy which limits how much video I can download... I would love to read it but that would mean a lot of work for someone or other (or a transcription software could be used perhaps)

 

yes, that plus something called a Fair Access Policy which limits how much video I can download... I would love to read it but that would mean a lot of work for someone or other (or a transcription software could be used perhaps)

Yes, I was hoping that there might be a transcription as well! But it would be a lot of work.

 

If you're going to watch, Alter is first, then Gill, then Lo, and then the Q & A. I recommend you skip Gill if you don't want to be spurred into instant activism, although he is also in the Q & A part. You can view the slides separate from the video at the same link.

 

Satterfield is considered an expert?

Satterfield finished his dissertation in December 2007. I assume he finished his PhD in 2008, the same year he started Cooperative Diagnostics. However, he gave what looks to be a grad student seminar at Arizona State University in 2009 ( http://www.bme.arizona.edu/Seminar/Schedules/seminar_fall09.php ) so his graduation date is unclear.

The extent of his work with United States Army Medical Research Institute of Infectious Disease, Department of Homeland Security, National Biodefense Analysis and Countermeasures Center is also unclear.

He was awarded a Department of Homeland Security DHS education award in 2005 ( http://www.biodesign.asu.edu/news/reaves-earns-prestigious-homeland-security-honors ) to pursue his graduate studies.

He did internships for one or more summers at Sandia National Laboratories.

Here is Cort's account of this:

Throughout his career he has been firmly ensconsed in the PCR arena and has developed numerous products to enhance PCR’s effectiveness. He is the founder, CEO and President of Cooperative Diagnostics , a medical diagnostic lab based in South Carolina.

Dr. Satterfield’s creation of new fast acting diagnostic probes with enhanced sensitivities which were able to withstand high temperatures apparently evoked interest from Homeland Security and he has worked with United States Army Medical Research Institute of Infectious Disease, Department of Homeland Security, National Biodefense Analysis and Countermeasures Center.

 

Satterfield is considered an expert?

Thanks for the info lulu...
Satterfield isn't interested in pursuing XMRV research so I'm going to focus my attention on those who are still doing XMRV research.

 

If you're going to watch, Alter is first, then Gill, then Lo, and then the Q & A. I recommend you skip Gill if you don't want to be spurred into instant activism, although he is also in the Q & A part. You can view the slides separate from the video at the same link.

Thanks gracenote...
I think I'll take your advice, and skip Gill to save myself some mental energy; I've had my fill of instant activism since the PACE trial was published!

 

Satterfield is considered an expert?

Satterfield finished his dissertation in December 2007. I assume he finished his PhD in 2008, the same year he started Cooperative Diagnostics. However, he gave what looks to be a grad student seminar at Arizona State University in 2009 ( http://www.bme.arizona.edu/Seminar/Schedules/seminar_fall09.php ) so his graduation date is unclear.

The extent of his work with United States Army Medical Research Institute of Infectious Disease, Department of Homeland Security, National Biodefense Analysis and Countermeasures Center is also unclear.

He was awarded a Department of Homeland Security DHS education award in 2005 ( http://www.biodesign.asu.edu/news/reaves-earns-prestigious-homeland-security-honors ) to pursue his graduate studies.

He did internships for one or more summers at Sandia National Laboratories.

If Satterfield does not have enough expertise to make comments regarding XMRV, how on earth can all you people from the other forum who keep posting here say that Gerwyn's comments hold any water? Doesn't he have a PhD in psychology -- Satterfield's comments do have a lot of merit because he does have some pretty relevant education and experience if you choose to look it up.

I think Satterfield has made some very good points. Trying to impugn the reputation of Satterfield does not make the comments any less true. The science will speak for itself.

Thanks Cort for this thread and the Satterfield interview.

 

Although Satterfield agrees with this, Judy Mikovits, Ian Lipkin, Harvey Alter and Frank Ruscetti do not. So we have experienced, award winning virologists at WPI, the Cleveland Clinic, the National Cancer Institute, and the NIH on the one hand, and a newly minted biomedical engineering PhD with a brand new start up company on the other.

Actually you have Abbott labs - with probably the premier antibody development platform in the country on the other side as well. They ended up spending $50,000 a piece on macaques to develop their antibody test because they didn't feel they had a validated positive patient sample.

They presumably know what they are doing. They created the first HIV antibody test. They are reportedly using their XMRV antibody test to do rapid testing of large numbers of blood samples now.

From James Curran, M.D., MPH, dean and professor of epidemiology of the Rollins School of Public Health, Emory University: "Abbott’s HIV antibody test was a pivotal public health milestone that ushered an era of scientific progress against AIDS, in which untold millions of lives were saved through enhanced detection of the virus and prevention of its transmission. In my opinion, this breakthrough was a landmark medical milestone for protecting public health. Blood wasn’t safe until the HIV antibody test arrived at our labs."

So we have experts on both sides.....no surprise there!

 

Thanks, Cort and Dr. Satterfield, for this interview.
I want to find out the truth about XMRV and its relationship to ME/CFS, of course.
It seems clear that Dr. Satterfield has the best of intentions and is well qualified in his field.

I just have the feeling that XRMV is going to present some kind of paradigm shift.
Even the smartest person and the most educated and competent person could miss some little detail that he didn't know mattered. Like refreezing a blood sample or etc.

 

http://videocast.nih.gov/Summary.asp?File=16477

Demystifying Medicine - Chronic Fatigue Syndrome: Is there a virus?

I've watched some of the video, and I've transcribed the most relevant section that I've watch so far.
Some of Lo's presentation is a bit disjointed, towards the end of the quote, below, but the only places that i've missed out any text is in places where i've placed 3 dots (...).
I've bolded the most relevant bits for this discussion.


Shyh-Ching Lo presentation:

[83.05]

"So why did many studies have different findings? This is obviously a very challenging question...

...


"...and the way the clinical sample is prepared, and the processing of this, all can make a difference.

[83.45]

And even more possible to me is that there is a variation of the PCR protocol, although everybody says we are following the same PCR assay, but if you look into all the detail, the cycles are different, [indiscernable] temperature slightly different, magnesium concentration slightly different;

all of this, we really don't know how much that's going to make a difference.

Today the topic is, is there a virus or not? is the virus responsible, or the causative agent of this, or not?

That's all very far away at the present time because, when we are looking at this, we obviously are dealing with a very low rate [or 'grade'?] of infection - a very low copy number in the blood,

and many of these difference can certainly result in the PCR disparities,

and i just want to mention... the NIH obviously, the NHLBI's, is looking into this, and have this sample, coded sample, sent to different laboratories, and to test it,

and this is the clinical [or 'critical'?] panel, the CDC's result,

and obviously ... the four of the patients,

and depends how the sample is being processed,

how long the delay of the processing of the sample, and the results are obviously different;

some are negative and some are positive,

so this is obviously, they also continue to look into this and try to solve is there any processing of the sample make a difference?

..."
[85.30]

http://videocast.nih.gov/Summary.asp?File=16477

Slide no. 93:

Why did many other studies have different findings--

There could be a difference in the prevalence of the viral agents among patient groups in different geographic areas.

Heterogeneity of CFS patient groups could be significant.

Variations of clinical sample preparations could affect PCR amplification effectiveness and assay sensitivity.

Variations of PCR protocols, primers, reagents or assay designs may have different sensitivity in detecting the diverse group of MLV-related virus gene sequences in the clinical samples.

The nature of low grade infections with low titers of the virus or low copy numbers of the viral target genes in patients' blood may likely account for the inconsistence and the PCR disparity.

 

Here's more from the video... This is interesting...

At [96.55]:

Harvey and Lo sent two of their positive patient samples to the CDC who then couldn't detect the PMRV's in the samples.

The CDC sent a blind coded negative control sample, from a single patient, to Harvey and Lo, which Harvey and Lo then repeatedly identified as positive, about 15 times. They repeatedly identified the same sample as being positive, even though it was blind coded. So Harvey and Lo detected PMRV in a 'negative control' from the CDC. Lo seems to be saying that this means it isn't a negative sample, but that the CDC just couldn't detect the virus in the sample.

[My thoughts: if we are talking about contamination here, then it couldn't be in the sample itself, but must enter during the testing stage. But Harvey and Lo's consistent and repeated testing of the CDC's blinded negative control sample, as positive, suggests that it isn't contamination.]

[99.10]

Lo: "We are pushing to the very end of the sensitivity..."

 

very interesting, thanks very much Bob.

 

Money interests

Thanks Cort, very positive and upbeat article, and even if our "favourite" virus is not quite there it may be part of the picture still.

I particularly appreciate how you pointed out at the beginning of the article how much Satterfield has invested in PCR. His entire career, livelihood and professional life is based around PCR. As he says, it's his profession.

It doesn't appear he's looking for XMRV as much as he's looking to find XMRV only through his style of PCR testing.

It's more like -- here's my method -- I can't find it -- there is no XMRV.

 

cort,

do we know anything about what abbott labs is doing with their antibody test in regard to humans?

 

I've done some more transcription of the Alter/Lo presentation, and posted it on an appropriate thread:

http://forums.aboutmecfs.org/showth...-Lo-and-Alter!&p=166303&viewfull=1#post166303

 

Here's more from the video... This is interesting...

At [96.55]:

Harvey and Lo sent two of their positive patient samples to the CDC who then couldn't detect the PMRV's in the samples.

The CDC sent a blind coded negative control sample, from a single patient, to Harvey and Lo, which Harvey and Lo then repeatedly identified as positive, about 15 times. They repeatedly identified the same sample as being positive, even though it was blind coded. So Harvey and Lo detected PMRV in a 'negative control' from the CDC. Lo seems to be saying that this means it isn't a negative sample, but that the CDC just couldn't detect the virus in the sample.

[My thoughts: if we are talking about contamination here, then it couldn't be in the sample itself, but must enter during the testing stage. But Harvey and Lo's consistent and repeated testing of the CDC's blinded negative control sample, as positive, suggests that it isn't contamination.]

[99.10]

Lo: "We are pushing to the very end of the sensitivity..."

All very interesting Bob - thanks for that

I think Satterfield thinks that the PCR community has covered the most of the bases or contingencies, I guess you would say, now -too many to expect that XMRV is there and they can't find it.......and Harvey and Lo clearly think they haven't and that there are still little differences that could have caused the virus to evade detection......

Of course there is XMRV AND the MLV's...lots of interesting stuff.

Mikovits was just at Univ of Alberta training them in how to find it in their new lab - so they are going to be important players in all this as well.

....the next couple months are going to be fascinating.

 

cort,

do we know anything about what abbott labs is doing with their antibody test in regard to humans?

Yes we do - they are using them for large-scale 'though-put' blood testing. I imagine this is why they spent the money on the macaques..I think its already started.

From CROI

X

MRV: Examination of Viral Kinetics, Tissue Tropism, and Serological Markers of Infection
X Qiu1, P Swanson1, K-C Luk1, J Das Gupta2, N Onlamoon3, R Silverman2, F Villinger3, S Devare1, G Schochetman1, and John Hackett, Jr*1

1Abbott Diagnostics, Abbott Park, IL, US; 2Cleveland Clin, OH, US; and 3Yerkes Natl Primate Res Ctr, Emory Univ, Atlanta, GA, US

Background: Xenotropic Murine Leukemia Virus-related Retrovirus (XMRV) is a human retrovirus recently discovered in familial prostate cancer tissue using DNA array based Virochip technology. Understanding viral replication kinetics, tissue tropism, and the host immune response is fundamental to establish the etiology of XMRV infection in human disease. Development of serologic assays to detect XMRV-specific antibodies would facilitate epidemiologic studies.

Methods: Five rhesus macaques were inoculated intravenously with XMRV. Blood was collected throughout the course of infection, and tissue from multiple organs was harvested at necropsy. Two macaques were necropsied at day 6 or 7 and one at day 144 post infection. The remaining 2 animals were re-inoculated with XMRV on day 158 and necropsied on day 291. XMRV-specific immunoreactivity was monitored by Western blot using viral lysate. Recombinant env gp70, p15E and gag p30 were utilized to develop serologic assays on the high-throughput automated ARCHITECT instrument system (Abbott Diagnostics).

Results: XMRV inoculation resulted in low transient plasma viremia, although proviral DNA persisted in circulating peripheral blood mononuclear cells for several weeks. Of interest, the earliest leukocyte targets were CD4+ T cells and NK cells followed by CD8+ enriched T and CD20+ enriched B cells (50% positive); CD14+ monocytes were negative. Animals sacrificed at the acute stage showed evidence of viral replication in spleen, lung, lymph nodes and liver. In contrast, sacrifice of 2 animals at 19 weeks post XMRV re-inoculation showed greater dissemination of XMRV DNA and RNA in various organs including the GI and urinary tract as well as in vaginal tissue of the one female.

By Western blot analysis, all 3 chronically infected macaques developed antibody responses to env and gag proteins. The serologic assays demonstrated 100% sensitivity by detecting all Western blot positive serial bleeds from the XMRV-infected macaques.

Preliminary results showed evidence of detectable reactivity to all 3 antigens in a low proportion (~0.1%) of US blood donors.

Conclusions: These data suggest that lymphocytes are a primary target for replication persistence (low grade replication) of XMRV in the absence of detectable plasma viremia. This study identified specific serological markers useful for detection of antibodies induced by XMRV infection. The prototype antibody assays will facilitate large-scale epidemiological studies.

 

I particularly appreciate how you pointed out at the beginning of the article how much Satterfield has invested in PCR. His entire career, livelihood and professional life is based around PCR. As he says, it's his profession.

It doesn't appear he's looking for XMRV as much as he's looking to find XMRV only through his style of PCR testing.

It's more like -- here's my method -- I can't find it -- there is no XMRV.

Yes with the proviso that he's talking about methods that most people use to search for viruses now -ie PCR and antibodies...I think you're right - he is heavily invested in these techniques, he believes they are the best techniques in his opinion and if you can't find it using them - then he doesn't believe they are there. I think that is what he is asserting.

 

I particularly appreciate how you pointed out at the beginning of the article how much Satterfield has invested in PCR. His entire career, livelihood and professional life is based around PCR. As he says, it's his profession.

It doesn't appear he's looking for XMRV as much as he's looking to find XMRV only through his style of PCR testing.

It's more like -- here's my method -- I can't find it -- there is no XMRV.

Brilliant observation Caroline!