Re:Lyme/genetics/ BowenTim

[Guest guest]

Tim:

Bowen is a controversial lab, as their method hasn't been able to be

replicated. The argument is that they don't properly identify what

they're taking micrographs of.

BARb

> >

> > a,

> >

> >

> > I should also mention that Jemsek said that I do not have the

> genetics to

> > be susceptible to Lyme.

> >

> >

> > Tim

>

[Guest guest]

> Tim:

> Bowen is a controversial lab, as their method hasn't been able to be

> replicated. The argument is that they don't properly identify what

> they're taking micrographs of.

>

> BARb

But who has ever tried to replicate their stuff? No one I know of.

I have never used primary immunofluoromicroscopy, but it is pretty

simple. I feel like one might have a fairly difficult time messing it

up.

I don't think the controversy over Bowen is worth a dime, at least as

far as I've been able to get ahold of it. Everyone I've read attacking

Bowen on lymenet, has *no* idea what they are talking about... (and I

emailed at least one of the most strident, to try to get down to brass

tacks about the basis for his belief.) There is also opposition from

certain UK and/or Euro health authorities, or there was 2 years ago

when I last paid attention to it. Those people may or may not have

defensible regulatory/procedural reasons for their opposition to the

use of the Bowen in diagnosis... I don't know how licensure of a

diagnostic test requires; there may a need for a peer-reviewed

publication or something, or not. Anyway, while I *don't* actually

know, I truly doubt that those regulatory people have made an inquiry

into the science that I would personally find acceptable... since after

all, virtually all the rest of the debate over lyme has ignored

concepts like a low-turnover pathogenic infection, or non-genomic abx

tolerance, which are probably key to explaining how a

(possible/probable) bacteriosis might be frequently seronegative and

respond to abx only extremely slowly.

On the other hand, I have seen statements coming out of Bowen regarding

the immunology of Bb infection and of CWD infection, that are hair-

raisingly... wrong, or at least wholly unsupported to the best of my

knowledge. So I don't trust them implicitly by any means. Far from

it... honestly, I am kind of leery. All I can say is that the test is

pretty simple, the procedure is well-described in their patent, and the

bodies in the micrographs I've seen do appear to be nicely fluoro-

labeled, at least to my inexperienced eye.

[Guest guest]

Exactly my point. Doesn't mean the bug is Lyme.

If they're finding Lyme in those blood samples.. i.e. if the bugs in

those pictures are Lyme.. then every one of those blood samples

should test PCR positive for Lyme. That ain't happenin'.

Barb

said:

All I can say is that the test is

pretty simple, the procedure is well-described in their patent, and

the

bodies in the micrographs I've seen do appear to be nicely fluoro-

labeled, at least to my inexperienced eye.

[Guest guest]

> Exactly my point. Doesn't mean the bug is Lyme.

I think it basically does mean it is lyme, though it is definitely

not one of the highest-confidence techniques. But polyspecific

antibody raised against a whole organism is pretty common to use in

research, I think. I am not certain, but I think that if you can

robustly label some microscopic body with an antibody against whole

organism " X " ... you can pretty much call that body an " X " in your

paper. If the labelling were quite weak, you would suspect a specious

cross-reaction of the antibody.

Certainly antibodies do cross-react. And when you use a polyspecific

antibody sample to label something, there are hundreds of different

antibody clones in there against different proteins... more chances

for cross-reactions. Labelling something with a monoclonal antibody

(a sample of antibody consisting purely of one antibody clone, which

only hits one protein of the organism) gives you more confidence.

However, it's harder to get robust labelling with a monoclonal,

because many proteins just aren't that abundant on the organism's

surface.

It would be /great/ if Bowen could light those bodies up with some

monoclonal antibodies against Bb. Do that with a couple different

monoclonals (with proper negative controls), and you're really beyond

all doubt. Why don't they publish something like this? I don't know.

Even if it is published in some two-bit journal that no one reads,

you can still cite it (in the event of controversy) and people can

still read it and judge for themselves... it's still science. And if

you can publish it in Infection and Immunity or Nature, so much the

better.

Notice they already do use an E coli negative control, and their

stuff doesn't label E coli at all. That's certainly a good sign.

Getting 20 different organisms and trying them all as negative

controls may be even better. Actually, Kirkegaard and , who

manufacture the antibody, may have already done that... it may be

standard procedure. I'm not sure. Their web catalog doesn't say

anything about that though, that I could find.

> If they're finding Lyme in those blood samples.. i.e. if the bugs

in

> those pictures are Lyme.. then every one of those blood samples

> should test PCR positive for Lyme. That ain't happenin'.

> Barb

That is the truth. What is going on there, I can't say. This

certainly does increase the odds that something weird is going on...

but I think there could also be some problem with the PCR. I am a

little shy of PCR myself. It usually seems to work well, but I do

notice instances where it gives weird results... for instance, many

many labs now agree that you can amplify C trach from the joint in C

trach ReA... but prior to that, a study had come out saying these

joints were PCR-negative.

I also have run PCRs in the lab, and had a hard time getting them to

work myself, quite often.

I must admit that it's easy to do a positive control PCR that is very

attractive intellectually: just add 10 borreliae to a microliter

tissue sample, and if it comes out positive, there ya go, your PCR

has a sensitivity of 10 borreliae / microliter, in that tissue -- and

there should be no room for excuses about some unknown PCR inhibitors

being in the samples.

Yet we still see these problems, like Stratton's lab shows Cpn by PCR

in these multiple sclerosis samples... some other lab finds nothing

in the same samples... so Stratton sent some other positive samples

he's done (or maybe it was the same samples from before; I forget) to

a third academic lab, which gets PCR results pretty close to the same

as Stratton's. A headache.

> said:

> All I can say is that the test is

> pretty simple, the procedure is well-described in their patent, and

> the

> bodies in the micrographs I've seen do appear to be nicely fluoro-

> labeled, at least to my inexperienced eye.