Re: Dura - bowen - question about specificity / different strains

[Guest guest]

As many people I have been wondering a lot about the bowen test. By

the way, personally I tested positive, 11 months ago 1:128 highest

serial dilution (wasn t on abx when the test wa carried out) and 2

weeks ago 1:16 serial dilution (was on abx when test was carried out),

I also improved somewhat (not spectacular) symptomswise, which migh

correlate with the lower dilution.

I looked at the patent and could not find anything about the borrelia

strains which are tested. Nothing is mentioned about sensu stricto or

lato, garinii, afzeli, lusitiana, valaisiana, etc. and I think I read

there are quite a few non pathogenic strains of borrelia.

Most stuff on this list is way over my head, and I do not have the

knowledge most you guys have to understand these things, but in

the below message seemns to mention hot specificity is guaranteed in

the bowen test, excluding hermsii for instance, so I assume it should

also be mentioned to which strains the QRIBB test is supposed to

respond? I do not see the cited text in the patent by the way.

I think some people on lymenet or other lists have written to bowen

asking which strains are testeb but i have never read an answer, does

anyone on this list know?

Adios,

> They use a polyclonal antibody. You get it from the blood of animals

> injected with Bb. Im not sure why the animals have abundant antibody

> when many infected humans do not. It may be because the exterior

> antigens are expressed more abundantly in vitro than in vivo, as some

> studies suggest. Injecting 10^7 lab-raised Bb might produce alot of

> antibody because they have alot of antigen.

>

> Im not sure how much this animal antiserum is purified, other than to

> say that one gets rid of all the blood cells of course. There may be a

> way to get rid of some serum proteins etc.

>

> Then, to ensure specificity, you then use affinity adsorbtion. This

> part is cool. You dont want your stuff to react with B hermsii, but

> the latter shares some of the same antigens as Bb, so it will bind

> some of the antibody youve raised against Bb. So you run a bunch of

> dead B hermsii thru your stuff till all the antibodies that bind to

> hermsii are gone.

>

> The antibody is then conjugated with a fluorescent molecule.

>

> You can look up the specs for the stuff ref'd in the Bowen patent:

>

> " Cross reactivity to Borrelia hermsii, Borrelia

> coriaceae, and Borrelia anserina has been minimzed

> through extensive affintiy adsorption. "

>

> " antibody has not be tested for

> cross-reactivity for treponemes "

>

> ...but if a cross-reaction reveals a bunch of treponemes in ones

> blood, that too could be certainly connected to ones ill health...

[Guest guest]

I wrote to Bowen in August last year & asked

" Does the Q-RIBb test show positive to species of borrelia other than

burgdorferi, and if so which ones "

The reply was

" The research test is extremely specific to Borrelia burgdorferi. "

Which doesn't distinguish between Bb sensu stricto & Bb sensu lato. So I

wrote back asking whether they meant Bb ss or Bb sl & had no reply.

Sue

panchitocarioca wrote:

> I looked at the patent and could not find anything about the borrelia

> strains which are tested. Nothing is mentioned about sensu stricto or

> lato, garinii, afzeli, lusitiana, valaisiana, etc. and I think I read

> there are quite a few non pathogenic strains of borrelia.

>

> I think some people on lymenet or other lists have written to bowen

> asking which strains are testeb but i have never read an answer, does

> anyone on this list know?

[Guest guest]

The three LYME sensu lato species are:

B burdorferi sensu stricto, B garinii and B afzelli.

Borrelia hermsii and B. turicatae transmit relapsing fever

(not Lyme) (although Alan Barbour has done some recent work

raising some questions in my mind if these are distinct).

And there are other species that are said not to be pathogenic -

so I won't list them.

Usually, depending on where the infected tick lives - it's infected

with one species (i.e. afzelli being European and burdorferi sensu

stricto being north american).. but researchers have found ticks

infected with more than one species in Europe. (In USA I think only

sensu stricto has been found in the ticks gut - but I'd have to

check).

To test for specific species- the primers (for PCR) would have to be

genospecific. You have to ask the lab that's running the PCR what

they're using- I think all the diagnostic PCRs use a universal

primer - so the species can't be determined. But I also know that a

universal primer can be tweaker toward being more or less specific to

the 3 species- should they want to do that.

But sometimes researchers test ticks with genospecific primes to

ID the species the tick harbors.

SInce researcher have shown people in Europe can be infected with

all three species- most labs for PCR - use the universal primer -

although I know that not all primers are the same- so what the

specificity is one can't say for sure.

I guess the question I'd have to Bowen (and I've never contacted

them) is what test are you using. I don't know if their test is a PCR

or not.

The pictures they take would mean nothing to me if I didn't know what

tests they're performing on the blood.

Barb

> > They use a polyclonal antibody. You get it from the blood of

animals

> > injected with Bb. Im not sure why the animals have abundant

antibody

> > when many infected humans do not. It may be because the exterior

> > antigens are expressed more abundantly in vitro than in vivo, as

some

> > studies suggest. Injecting 10^7 lab-raised Bb might produce alot

of

> > antibody because they have alot of antigen.

> >

> > Im not sure how much this animal antiserum is purified, other

than to

> > say that one gets rid of all the blood cells of course. There may

be a

> > way to get rid of some serum proteins etc.

> >

> > Then, to ensure specificity, you then use affinity adsorbtion.

This

> > part is cool. You dont want your stuff to react with B hermsii,

but

> > the latter shares some of the same antigens as Bb, so it will bind

> > some of the antibody youve raised against Bb. So you run a bunch

of

> > dead B hermsii thru your stuff till all the antibodies that bind

to

> > hermsii are gone.

> >

> > The antibody is then conjugated with a fluorescent molecule.

> >

> > You can look up the specs for the stuff ref'd in the Bowen patent:

> >

> > " Cross reactivity to Borrelia hermsii, Borrelia

> > coriaceae, and Borrelia anserina has been minimzed

> > through extensive affintiy adsorption. "

> >

> > " antibody has not be tested for

> > cross-reactivity for treponemes "

> >

> > ...but if a cross-reaction reveals a bunch of treponemes in ones

> > blood, that too could be certainly connected to ones ill health...

[Guest guest]

My additional question is not about the strain but rather about the

specificity... clearly the sensitivity is there (100%????), but my

concern is with the specificity... not only of the Bb probe (and

presumably the HGE, bart, bab), but also of the florescent aspect...

> > They use a polyclonal antibody. You get it from the blood of

animals

> > injected with Bb. Im not sure why the animals have abundant

antibody

> > when many infected humans do not. It may be because the exterior

> > antigens are expressed more abundantly in vitro than in vivo, as

some

> > studies suggest. Injecting 10^7 lab-raised Bb might produce alot

of

> > antibody because they have alot of antigen.

> >

> > Im not sure how much this animal antiserum is purified, other

than to

> > say that one gets rid of all the blood cells of course. There

may be a

> > way to get rid of some serum proteins etc.

> >

> > Then, to ensure specificity, you then use affinity adsorbtion.

This

> > part is cool. You dont want your stuff to react with B hermsii,

but

> > the latter shares some of the same antigens as Bb, so it will

bind

> > some of the antibody youve raised against Bb. So you run a bunch

of

> > dead B hermsii thru your stuff till all the antibodies that bind

to

> > hermsii are gone.

> >

> > The antibody is then conjugated with a fluorescent molecule.

> >

> > You can look up the specs for the stuff ref'd in the Bowen

patent:

> >

> > " Cross reactivity to Borrelia hermsii, Borrelia

> > coriaceae, and Borrelia anserina has been minimzed

> > through extensive affintiy adsorption. "

> >

> > " antibody has not be tested for

> > cross-reactivity for treponemes "

> >

> > ...but if a cross-reaction reveals a bunch of treponemes in ones

> > blood, that too could be certainly connected to ones ill

health...

[Guest guest]

Its an antibody-antigen thing. The antibody is raised in animals

against some particular Bb, and the antibody is polyclonal meaning it

has a bunch of differently-shaped antibodies recognizing a bunch of

different antigens. A tiny change in an antigen will prevent it from

binding the entibody, but closely-related species will I think

generally share some common antigens that are precisely alike.

My guess would be that RIBb would be sensitive to all Bb sensu lato

but thats just a guess. To be certain, youd need to find evidence of

what their " antigenic " or " serological " relationship is. Over at

eurolyme they use the RIBb alot, and I believe they would be

very concerned with the possibility of Bbs other than sensu stricto -

so they could probably be more helpful. Perhaps theyve discussed it

in the past.

Then theres B lonestari, which I know about only from soft sources...

I guess its outside Bb sensu lato altogether and I dont know what its

serological relationship would be.

Its also possible that distinct serotypes could be involved in non-

lyme diseases like ALS, MS, Alzheimers, if they are spirochetal.

To take one concrete example, the antibody used by the Brorson study

of MS " was produced against B. burgdorferi, but is also known to

react with other spirochetes (Treponema pallidum, Borrelia hermsii,

and Borrelia parkerii). " This antibody is evidently *not* affinity

adsorbed as the Bowen stuff is.

" panchitocarioca " <r_steentjes@h...> wrote:

>

> As many people I have been wondering a lot about the bowen test. By

> the way, personally I tested positive, 11 months ago 1:128 highest

> serial dilution (wasn t on abx when the test wa carried out) and 2

> weeks ago 1:16 serial dilution (was on abx when test was carried

out),

> I also improved somewhat (not spectacular) symptomswise, which migh

> correlate with the lower dilution.

>

> I looked at the patent and could not find anything about the

borrelia

> strains which are tested. Nothing is mentioned about sensu stricto

or

> lato, garinii, afzeli, lusitiana, valaisiana, etc. and I think I

read

> there are quite a few non pathogenic strains of borrelia.

>

> Most stuff on this list is way over my head, and I do not have the

> knowledge most you guys have to understand these things, but in

> the below message seemns to mention hot specificity is guaranteed in

> the bowen test, excluding hermsii for instance, so I assume it

should

> also be mentioned to which strains the QRIBB test is supposed to

> respond? I do not see the cited text in the patent by the way.

>

> I think some people on lymenet or other lists have written to bowen

> asking which strains are testeb but i have never read an answer,

does

> anyone on this list know?

>

> Adios,

>

>

>

>

>

>

> > They use a polyclonal antibody. You get it from the blood of

animals

> > injected with Bb. Im not sure why the animals have abundant

antibody

> > when many infected humans do not. It may be because the exterior

> > antigens are expressed more abundantly in vitro than in vivo, as

some

> > studies suggest. Injecting 10^7 lab-raised Bb might produce alot

of

> > antibody because they have alot of antigen.

> >

> > Im not sure how much this animal antiserum is purified, other

than to

> > say that one gets rid of all the blood cells of course. There may

be a

> > way to get rid of some serum proteins etc.

> >

> > Then, to ensure specificity, you then use affinity adsorbtion.

This

> > part is cool. You dont want your stuff to react with B hermsii,

but

> > the latter shares some of the same antigens as Bb, so it will bind

> > some of the antibody youve raised against Bb. So you run a bunch

of

> > dead B hermsii thru your stuff till all the antibodies that bind

to

> > hermsii are gone.

> >

> > The antibody is then conjugated with a fluorescent molecule.

> >

> > You can look up the specs for the stuff ref'd in the Bowen patent:

> >

> > " Cross reactivity to Borrelia hermsii, Borrelia

> > coriaceae, and Borrelia anserina has been minimzed

> > through extensive affintiy adsorption. "

> >

> > " antibody has not be tested for

> > cross-reactivity for treponemes "

> >

> > ...but if a cross-reaction reveals a bunch of treponemes in ones

> > blood, that too could be certainly connected to ones ill health...

[Guest guest]

" Then theres B lonestari, which I know about only from soft

sources...

I guess its outside Bb sensu lato altogether and I dont know what its

serological relationship would be. "

Doesn't Igenex claim their test is sensitive for lonestari? And

hasn't the CDC conceded it does indeed appear to be causing Lyme?

> > > They use a polyclonal antibody. You get it from the blood of

> animals

> > > injected with Bb. Im not sure why the animals have abundant

> antibody

> > > when many infected humans do not. It may be because the

exterior

> > > antigens are expressed more abundantly in vitro than in vivo,

as

> some

> > > studies suggest. Injecting 10^7 lab-raised Bb might produce

alot

> of

> > > antibody because they have alot of antigen.

> > >

> > > Im not sure how much this animal antiserum is purified, other

> than to

> > > say that one gets rid of all the blood cells of course. There

may

> be a

> > > way to get rid of some serum proteins etc.

> > >

> > > Then, to ensure specificity, you then use affinity adsorbtion.

> This

> > > part is cool. You dont want your stuff to react with B

hermsii,

> but

> > > the latter shares some of the same antigens as Bb, so it will

bind

> > > some of the antibody youve raised against Bb. So you run a

bunch

> of

> > > dead B hermsii thru your stuff till all the antibodies that

bind

> to

> > > hermsii are gone.

> > >

> > > The antibody is then conjugated with a fluorescent molecule.

> > >

> > > You can look up the specs for the stuff ref'd in the Bowen

patent:

> > >

> > > " Cross reactivity to Borrelia hermsii, Borrelia

> > > coriaceae, and Borrelia anserina has been minimzed

> > > through extensive affintiy adsorption. "

> > >

> > > " antibody has not be tested for

> > > cross-reactivity for treponemes "

> > >

> > > ...but if a cross-reaction reveals a bunch of treponemes in

ones

> > > blood, that too could be certainly connected to ones ill

health...

[Guest guest]

:

You and others need to understand how these are classified I guess.

Borrelia is classified as the GENUS

There are over 15 or 20 SPECIES of the GENUS borrelia -

Very few of these species are known to cause disease in humans (but

they're finding more everyday it seems).

TWO of the SPECIES GROUPS of the GENUS borrelia transmit disease to

humans are for Lyme (tick tramsitted) and relapsing fever (flea

transmitted)

LoneStari was originally classified with the Lyme causing group, but

has been re-classified, based on gene sequencing, and oput into the

species group that casues Relapsing Fever.

When testing - Specificity get's harder in the following descending

order.. Genus, Species, then strain.

If Bowen doesn't classify even the Genus of the Spirochete- then it

lacks specificity - meaning it's not very speficic even to the

GENUS.. if this is the case- then no wonder why their results are

questioned..

Hope this answer clears up some confusion.

Barb

> > > > They use a polyclonal antibody. You get it from the blood of

> > animals

> > > > injected with Bb. Im not sure why the animals have abundant

> > antibody

> > > > when many infected humans do not. It may be because the

> exterior

> > > > antigens are expressed more abundantly in vitro than in vivo,

> as

> > some

> > > > studies suggest. Injecting 10^7 lab-raised Bb might produce

> alot

> > of

> > > > antibody because they have alot of antigen.

> > > >

> > > > Im not sure how much this animal antiserum is purified, other

> > than to

> > > > say that one gets rid of all the blood cells of course. There

> may

> > be a

> > > > way to get rid of some serum proteins etc.

> > > >

> > > > Then, to ensure specificity, you then use affinity

adsorbtion.

> > This

> > > > part is cool. You dont want your stuff to react with B

> hermsii,

> > but

> > > > the latter shares some of the same antigens as Bb, so it will

> bind

> > > > some of the antibody youve raised against Bb. So you run a

> bunch

> > of

> > > > dead B hermsii thru your stuff till all the antibodies that

> bind

> > to

> > > > hermsii are gone.

> > > >

> > > > The antibody is then conjugated with a fluorescent molecule.

> > > >

> > > > You can look up the specs for the stuff ref'd in the Bowen

> patent:

> > > >

> > > > " Cross reactivity to Borrelia hermsii, Borrelia

> > > > coriaceae, and Borrelia anserina has been minimzed

> > > > through extensive affintiy adsorption. "

> > > >

> > > > " antibody has not be tested for

> > > > cross-reactivity for treponemes "

> > > >

> > > > ...but if a cross-reaction reveals a bunch of treponemes in

> ones

> > > > blood, that too could be certainly connected to ones ill

> health...

[Guest guest]

> If Bowen doesn't classify even the Genus of the Spirochete- then it

> lacks specificity - meaning it's not very speficic even to the

> GENUS.. if this is the case- then no wonder why their results are

> questioned..

But - is that the case? I believe they name in their patent 2 sources

of direct immunofluoresence antibody, one of which is affinity

adsorbed with only non-burgdorferi-sensu-lato borreliae. They note it

is not adsorbed with any treponemes, so one could have

cross-reactivity with Treponema. One could also have cross-reactivity

with other non-burgdorferi-s-l species other than those used in the

affinity adsorbtion. But it might be limited - for instance if

Treponema were indeed cross-labelled by the RIBb and the labelling

were, say, 5x weaker than with Bbsl, then a technician with any

experience would recognize this immediately.

Anyway, if youre sick and your blood is full of forms of some kind of

poorly-abx-responsive spirochete, its news.

But, I absolutely agree, they should be running RIBb on every sort of

positive and negative control sample they can find, and publishing on

it extensively. Perhaps theres some reason why they arent, but I dont

know it. As it is they seem to be just going for FDA approval instead,

according to what I hear/rmemeber.

[Guest guest]

Thanks BP. VERY clear explanation!!

> > > > > They use a polyclonal antibody. You get it from the blood

of

> > > animals

> > > > > injected with Bb. Im not sure why the animals have

abundant

> > > antibody

> > > > > when many infected humans do not. It may be because the

> > exterior

> > > > > antigens are expressed more abundantly in vitro than in

vivo,

> > as

> > > some

> > > > > studies suggest. Injecting 10^7 lab-raised Bb might

produce

> > alot

> > > of

> > > > > antibody because they have alot of antigen.

> > > > >

> > > > > Im not sure how much this animal antiserum is purified,

other

> > > than to

> > > > > say that one gets rid of all the blood cells of course.

There

> > may

> > > be a

> > > > > way to get rid of some serum proteins etc.

> > > > >

> > > > > Then, to ensure specificity, you then use affinity

> adsorbtion.

> > > This

> > > > > part is cool. You dont want your stuff to react with B

> > hermsii,

> > > but

> > > > > the latter shares some of the same antigens as Bb, so it

will

> > bind

> > > > > some of the antibody youve raised against Bb. So you run a

> > bunch

> > > of

> > > > > dead B hermsii thru your stuff till all the antibodies

that

> > bind

> > > to

> > > > > hermsii are gone.

> > > > >

> > > > > The antibody is then conjugated with a fluorescent

molecule.

> > > > >

> > > > > You can look up the specs for the stuff ref'd in the Bowen

> > patent:

> > > > >

> > > > > " Cross reactivity to Borrelia hermsii, Borrelia

> > > > > coriaceae, and Borrelia anserina has been minimzed

> > > > > through extensive affintiy adsorption. "

> > > > >

> > > > > " antibody has not be tested for

> > > > > cross-reactivity for treponemes "

> > > > >

> > > > > ...but if a cross-reaction reveals a bunch of treponemes

in

> > ones

> > > > > blood, that too could be certainly connected to ones ill

> > health...

[Guest guest]

I have not looked into how Bowen does their testing.

I'd really have to do that indepth to make a comment or form an

opinion.

Barb

>

> > If Bowen doesn't classify even the Genus of the Spirochete- then

it

> > lacks specificity - meaning it's not very speficic even to the

> > GENUS.. if this is the case- then no wonder why their results are

> > questioned..

>

> But - is that the case? I believe they name in their patent 2

sources

> of direct immunofluoresence antibody, one of which is affinity

> adsorbed with only non-burgdorferi-sensu-lato borreliae. They note

it

> is not adsorbed with any treponemes, so one could have

> cross-reactivity with Treponema. One could also have cross-

reactivity

> with other non-burgdorferi-s-l species other than those used in the

> affinity adsorbtion. But it might be limited - for instance if

> Treponema were indeed cross-labelled by the RIBb and the labelling

> were, say, 5x weaker than with Bbsl, then a technician with any

> experience would recognize this immediately.

>

> Anyway, if youre sick and your blood is full of forms of some kind

of

> poorly-abx-responsive spirochete, its news.

>

> But, I absolutely agree, they should be running RIBb on every sort

of

> positive and negative control sample they can find, and publishing

on

> it extensively. Perhaps theres some reason why they arent, but I

dont

> know it. As it is they seem to be just going for FDA approval

instead,

> according to what I hear/rmemeber.