The Curious Case Of The IUML And The EPO Positive That Wasn't

[fmk_RoI]

Back in 2001, the IUML was at the forefront of the implementation of the then new EPO test. Things didn't always go quite as they should have: http://www.podiumcafe.com/2011/6/6/...e-of-the-iuml-and-the-epo-positive-that-wasnt

 

[thehog]

Back in 2001, the IUML was at the forefront of the implementation of the then new EPO test. Things didn't always go quite as they should have: http://www.podiumcafe.com/2011/6/6/...e-of-the-iuml-and-the-epo-positive-that-wasnt

Good article. Thank-you.

Saugy at it again. Will he ever stop?

--

In more recent months, the IUML has been back in the news concerning the EPO test and the early months of its implementation. Martial Saugy had this to say to the Neue Züricher Zeitung:

"They [the four suspect samples] were taken at four different stages [during the Tour de Suisse in June 2001], so I don't know whether they were from four different riders or all of the same athlete. But the tests were not covered up, and it is also not correct that they could have been interpreted as positive. They were suspect, and you wouldn't stand a chance at all with that sole argument in front of a court.

"The Paris laboratory of Chatenay-Malabry fixed the criteria for a positive test result. An athlete was positive only if eighty percent of the signs typical for the use of synthetic EPO were found. [A suspect sample was one that] showed between seventy and eighty percent of the typical EPO parameters. That meant that the probability of doping was high. But because such a result can also be produced naturally, it was all about excluding false positives."

Remember, this is Saugy speaking in 2011, nine years after the CAS pointed out that the UCI had no EPO threshold in 2001. Ten years after he and his IUML colleagues tried to bust Bo Hamburger using test results which fell below the 'threshold.' This is the same Martial Saugy who told the CAS then that:

"there was no scientific justification for an 80% threshold and that a level marginally below 80% was reliable enough to allow one to assume a positive result."

Is it any wonder that the IUML is sometimes referred to as being the UCI's lab in Lausanne?

 

[Master50]

Good article. Thank-you.

Saugy at it again. Will he ever stop?

Is it any wonder that the IUML is sometimes referred to as being the UCI's lab in Lausanne?

The UCI does not have a laboratory. They pay to have tests done at accredited labs that are not owned or operated by the UCI.
The Labs are not accredited to any UCI standard but are accredited to recognized international standards for anti doping testing. I think that is true of WADA who may set the standards for anti doping but also don't own the labs.

 

[fmk_RoI]

The UCI does not have a laboratory. They pay to have tests done at accredited labs that are not owned or operated by the UCI.
The Labs are not accredited to any UCI standard but are accredited to recognized international standards for anti doping testing. I think that is true of WADA who may set the standards for anti doping but also don't own the labs.

I didn't say the lab was the UCI's lab. I said it was no wonder it is referred to as being the UCI's lab.

As for UCI accreditation - I think you need to re-educate yourself.

 

[rata de sentina]

As I've said before, they should still have these gel images on file. The current method of analysing these images is less prone to false negatives than the old method. They don't need the sample just the image from the ccd camera of the gel. It's been shown that using different parameters and computerised analysis you can improve the detection rate, the sensitivity and the window following dosing.

The issue is that it is a subjective assay and the 80% criteria is just a crude attempt to quantitate it. At the end of the day CAS might not have been comfortable with that subjectivity but people who look at these things all the time will know "doped to the gills" when they see it on the gel. Subsequently of course the analysis has been refined because scientists were sick of looking at gels of people who were clearly doping but getting off.

Anyway, I can't beleive they are still stuffing about with this stupid IEF test. They should just subject the whole back catalog from the freezer to mass spec and go to town on them. They need to get some proper protein chemists on the job and stop fluffing about.

 

[Broken Spoke]

In close co-operation with other federations, the UCI validated the new test on 1 April 2001.

Oh dear...

 

[Alpe d'Huez]

Interesting thread, guys.

 

[DirtyWorks]

Required Reading

The linked article is as important as the 60 Minutes story. There's actually at least two full stories in the article. One about the efficacy of EPO testing and how the test protocol is used to suppress positives. The other about how the UCI enables doping.

---Topic 1
For those that have forgotten their statistics courses, here's a rough summary:

-No published false negative metric for the 2001-era test. That basically invalidates the test method because no one knows when a negative test is wrong.

-In 2007, (published 2008) a study testing the effectiveness of EPO testing by actually EPO doping subjects for seven weeks. Samples sent to WADA-accredited labs with horrific false negative results over 6 of the 7 weeks. Awful.

IOC's 2000 and 2002 test protocol of combining blood and urine results ensured an absolute minimum of positives because the likelihood of coming up positive on both is incredibly small.

---Topic 2
The article then goes on to discuss how the UCI enables doping. Federation mechanics is boring, but the article has fantastic information.

Hello, NY Times? Wall Street Journal? Here's a couple of stories that need retelling to a broader audience. The IOC's role in enabling doping is sloooowly coming to the foreground.

 

[biokemguy]

They need to get some proper protein chemists on the job and stop fluffing about.

+1

Sometimes it's pretty painful to look at their methods for catching dopers.

Unfortunately "proper protein chemists" cost money. We can make more than pro pack-fill by spending our time developing bigger d!(k drugs.

 

[Merckx index]

As I've said before, they should still have these gel images on file. The current method of analysing these images is less prone to false negatives than the old method. They don't need the sample just the image from the ccd camera of the gel. It's been shown that using different parameters and computerised analysis you can improve the detection rate, the sensitivity and the window following dosing.

The issue is that it is a subjective assay and the 80% criteria is just a crude attempt to quantitate it. At the end of the day CAS might not have been comfortable with that subjectivity but people who look at these things all the time will know "doped to the gills" when they see it on the gel. Subsequently of course the analysis has been refined because scientists were sick of looking at gels of people who were clearly doping but getting off.

Actually, I think they knew "doped to the gills" under the old system, too, but the linked article in the OP makes it clear what the problem is (the same problem with all doping tests): in order to minimize false positives, a standard is used that is certain to allow many false negatives. The new method might be better, it might show that a suspicious result back then is a clear positive today, but I'm sure it still includes many false negatives. Any test is going to have a substantial overlap between a typical doping profile and the far upper end of normal physiology. We all know that EPO use in the peloton is common, yet there are still very few busts.

Anyway, I can't beleive they are still stuffing about with this stupid IEF test. They should just subject the whole back catalog from the freezer to mass spec and go to town on them. They need to get some proper protein chemists on the job and stop fluffing about.

Ayotte is apparently working on this:

J Mass Spectrom. 2008 Jul;43(7):924-35.
Low LC-MS/MS detection of glycopeptides released from pmol levels of recombinant erythropoietin using nanoflow HPLC-chip electrospray ionization.
Groleau PE, Desharnais P, Coté L, Ayotte C.

I wonder, though, if it might not be necessary to use mass spec. According to this reference:

Blood. 2001 Dec 15;98(13):3626-34.
Sugar profiling proves that human serum erythropoietin differs from recombinant human erythropoietin.
Skibeli V, Nissen-Lie G, Torjesen P.

the biggest difference between rhEPO and natural EPO is that the synthetic protein has more sialic acid groups. Perhaps you could just first purify EPO from urine (which also would be necessary in an approach involving mass spec), remove sialic acid enzymatically, then measure the ratio of it to total EPO protein. Seems to me that this would be a lot faster and easier than mass spec, and avoid some of the complications of the latter.

However, either of these approaches is going to suffer from the same major problem that bedevils the current gel method. If the ratio of natural to synthetic EPO is high in a urine sample, it’s difficult to detect the key differences between the two molecules. Interestingly, I discovered that back in 2003 someone applied for a patent for a monoclonal antibody that was claimed to be specific for rhEPO. That would be the gold standard if possible, though of course you would need a different Ab for each new form of synthetic EPO. I don’t know what became of this procedure, though.

 

[biokemguy]

The monoclonal would be a gold standard and I've wondered why it hasn't been developed, but other molecules could be developed to bind rhEPO (any flavor).

RNA aptamers could work, or Darpins. Both would require time and investment but could ultimately be built into tests that would determine whether rhEPO was present or not...no ratio or threshold necessary.

 

[rata de sentina]

We shouldn't fall into the trap of merely considering rhEPO as one entity because some of the IEF patterns are quite different from different cells.

As I understand it rhEPO from CHO cells has carbohydrate moeties which are not found in natural epo so I still like mass spec. Protein chemists might be expensive but the current IEF method is also very pricey. Making a test more high throughput and more sensitive would save money in the long run. Mass specs are expensive to buy but not so hard to run and you can bang a lot of samples through them.