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» LymeNet Flash » Questions and Discussion » Medical Questions » WB Interpretation Help

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Author Topic: WB Interpretation Help
Jules17
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Yup, I am back. Here is what IgeneX and Dr. C came up with for me:

IgM:
Positive bands: 18, 28, 30, 31, 41 and 66.
18, 31 and 41 were high positives, if that matters.

Equivocal bands: 23-25, 34, 39, 45, 58 and 93.

So, IgeneX says my IgM is positive. My Dr. (Dr. C in MO) says to count equivocals as positives as there was something there.

On my IgG, however, IgeneX says negative.

Positive bands: 28 and 41.

Equivocal bands: 18, 31, 34, 39, 45, 58 and 66.

I am confused because I thought the IgM showed recent activity and the IgG was if you had been infected longer ago?

I was bitten 3 years ago so I am confused by that, but I am still in the LD embryo stage, knowledge-wise.

Thanks much for any interpretation help! Julie


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Carol in PA
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Julie,
Here is some info:

Interpreting the IgG & IgM Western Blot For Lyme Disease
Melissa Kaplan http://www.sonic.net/~melissk/wb.html

And The Bands Played On - Western blot serological test for Lyme disease http://www.geocities.com/HotSprings/Oasis/6455/western-blot.txt

Topic: Explaining Borreliosis (Lyme) Western Blot Tests http://flash.lymenet.org/ubb/Forum1/HTML/022767.html

Hope this helps!
Carol


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cootiegirl
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I pulled this out of one of the articles and thought this might help.....

With most infections, your immune system first forms IgM antibodies, then in about 2 to 4 weeks, you see IgG antibodies. In some infections, IgG antibodies may be detectable for years. Because Borrelia burgdorferi is a chronic persistent infection that may last for decades, you would think patients with chronic symptoms would have positive IgG Western blots.

But actually, more IgM blots are positive in chronic borreliosis than IgG. Every time Borrelia burgdorferi reproduces itself, it may stimulate the immune system to form new IgM antibodies. Some patients have both IgG and IgM blots positive. But if either the IgG or IgM blot is positive, overall it is a positive result

cootiegirl


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Lyddie
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Just to agree with the above. Positive IgM can indicate recent infection (most mainstream doctors interpret this way). But it can also indicate ongoing infection that has been around a long time, as cootie girl described.

Our family doctor continues to insist that 4 members of my family are getting bitten by ticks imediately before our respective Western Blots (what a coincidence!), and that that's the only possible reason for our positive IgM's. When I tell him that our feet never leave blacktop, he says that mice must be bringing ticks in our house. We've never seen a tick in our house.

BTW I had testing w/ a lot of equivocals at first. After a few weeks of antibiotics, I had even more IgM positives, more than the CDC requires for diagnosis.


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breathwork
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I recently attended a lecture by Dr. H in No CA.

He said that IgM persists throughout the course of the disease, can indicate more active lyme infection...

IgG merely indicates exposure, does not indicate if the bacteria are still present.

He also said that bands 31 and 34 are the most strongly indicative bands, but also mentioned 18, 23-25, 28, 39, 93, 58, and 83 as positive for lyme.


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Jules17
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So would I be considered positive???
Posts: 23 | From Lee's Summit, MO, USA | Registered: Sep 2004  |  IP: Logged | Report this post to a Moderator
rosesisland2000
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quote:
Originally posted by Jules17:
So would I be considered positive???

I don't understand your question???? YEs, you definitely would be considered positive, even if one of the IG's is negative.

IMO, you have more positives than I've seen in a long time. Don't put to much in store of which is positive or not. If you are positive on one, then you are positive.

Maybe you've not come across this in Dr. C's handouts yet. So here's what you have in your handouts that Dr. C gave you...his explaination of the WB tests.


Explaining Borreliosis (Lyme) Western Blot Tests
There is no universal agreement on what defines a positive Western blot.
Good laboratories use different criteria to interpret borreliosis blots. At the 1999 international borreliosis
and tick-borne infection conference, Sam Donta, M.D. lectured. Dr. Donta is a full professor of Infectious
Disease at Boston University School of Medicine. He said that if a patient has just one
borreliosis-associated antibody on their Western blot, you may assume they have borreliosis. Richard
Horowitz, M.D. said the same thing in his lecture, at that same conference.
Research I presented in 1998 involving over 400 borreliosis patients, showed an 87% response rate to
antibiotics. This was if they had one borreliosis-associated antibody on their blot. So if there is enough
suspicion that Lyme borreliosis is the cause of a patient's symptoms, so much so that a Western blot is
ordered, then if only one borreliosis-associated antibody is found, it is significant!
Medical literature is replete with statements about false positive test results for Lyme borreliosis. Since
1988, I have diagnosed and treated well over 600 borreliosis patients. Only 2 of those patients with a
positive borreliosis test did not respond to antibiotics. This is a 99% success rate!
So in the trenches of day-to-day medical practice, false positive borreliosis tests are not an issue. In
retrospect, those 2 patients that did not respond to antibiotics may have also had babesiosis. In my
practice, many borreliosis patients also have babesiosis, another tick-borne infection that causes the same
symptoms as Lyme borreliosis.
Babesiosis is caused by a protozoa, which is a different germ type than a bacteria, virus, fungus or yeast.
The placebo effect would not explain a 99% response rate. Those borreliosis associated antibodies should
not be there, in patients with symptoms.
A placebo is like a sugar pill, that has no effect. A placebo effect occurs because patients believe in the pill
they are taking, even though it is a sugar pill. The human mind causes the response. Placebo effects
should more likely be about 20-30%, not a 99% response rate.
False negative test results are the real problem in diagnosing borreliosis. Research has shown that you
have to do the right test (the Western blot), done at the right laboratory (one that specializes in testing
borreliosis), and done the correct way (shipped express delivery early in the week). The right test to screen
for borreliosis is the Western blot. Research I presented in Bologna, Italy in 1994 at the international
borreliosis conference showed this.
Other screening tests, such as the IFA, EIA, ELISA, and PCR DNA probe were often negative when the
Western Blot was positive! Other doctors like myself who diagnose and treat a lot of borreliosis patients,
go straight to the Western blot as their screening test.
Medical articles abound stating that it is best to do a screening test, such as an ELISA, and if it is positive,
then confirm it with a Western blot. But the ELISA is often negative when the Western blot is positive so,
the right test is the Western blot.
*** It lets you see exactly which antibodies are present. The "right laboratory" means one that specializes
in borreliosis testing. In the past, I have done head to head comparisons with 3 different regular labs.
Western blots were drawn and sent on the same day to 2 different labs. The labs that specialize in
borreliosis testing typically found borrelia-associated antibodies, that the regular laboratories missed. If
these specialty labs find a borrelia antibody, I trust it to be significant, because patients respond to
antibiotics.
**You get what you pay for, so use a lab that specializes in borreliosis. The right way to process the
Western blot specimen means for the blood to be drawn and express mailed early in the week. Research
shows the borrelia antibodies have the potential to clump together, resulting in false negative test results.
So far, unclumping has not been practical for laboratories to do.
The fresher the specimen, the more accurate the test results. Patients at our office are scheduled Monday,
Tuesday, or Wednesday if testing is to be done. This way, express shipping will assure that the specimen
does not spend the weekend sitting at the post office. This is the right way to test and ship borreliosis
specimens.
Western blots look for antibodies. These antibodies are made by your immune system. In this case, the
antibodies are made to fight against different parts of the Lyme bacteria, which is called Borrelia
burgdorferi, and other Borrelia species. In other words, your immune system does not make one big
antibody against the whole bacteria. So, when you see a number on a borreliosis Western blot, it
corresponds to a specific part of the bacteria.
Compare it to the old story of different blind people touching an elephant. Based on the part of the
elephant each one touched, each person had their own perception. Likewise, the antibodies attach to
different and specific parts of Borrelia burgdorferi.
Numbers on Western blots correspond to weights. Kilodaltons (kDa) are the units used for these
microscopic weights. Think of it like pounds or ounces. An 18 kDa antibody weighs 18 kilodaltons. To do
a Western blot, thin gel strips are impregnated with the various parts of Borrelia burgdorferi. Each of the
numbers, 18 through 93, on the test result form, is a part of the bacteria.
Blood is made up of red blood cells and serum; Spinning blood in a centrifuge separates serum from red
blood cells and other things, like white blood cells and platelets.
Serum contains antibodies made by the immune system. Electricity is used to push the serum through the
thin gel strips for the Western blot. If there are any antibodies against parts of Borrelia burgdorferi present
in your serum, and these parts are impregnated on the strip, the antibody will complex (bind) to that part.
When antibodies form a complex, it is called an antigen-antibody complex. Anything foreign in the body
is an antigen, such as a ragweed pollen particle, germ, cancer, and even a splinter.
In the case of borreliosis, the various parts of Borrelia burgdorferi are all antigens. Though each antigen is
different, they all come from the same bacteria. So all the numbers that are positive on the test report are
due to antigen-antibody complexes.
If enough of the complexes are formed, eventually it may be seen with the naked eye as a dark band. -
Band intensity reflects how dark or wide it is. Controversy exists about band intensity. Many would say
the " +/-" equivocal bands are not significant. The problem I have with that, is that there are "-" negative
bands. The lab has no trouble calling some bands negative. So they must be seeing something when they
put "+/-" at some bands.
The only thing that makes sense, is that there is a little bit of that antibody present in your serum. If the
"+/-" equivocal is reported on the borrelia associated bands, it is usually significant, in my clinical
experience. This is a strong clue that I am on the right track.
Instead of ignoring these, they should be a red flag to keep pursuing a laboratory diagnosis. Giving
patients 4 weeks of antibiotics (usually tetracycline, 500 mg, 3 times a day), will convert a negative or
equivocal Western blot to positive in about 36% of cases.
As mentioned, if these positive blots are found by specialty labs, over 99% of those patients will respond
to antibiotics.
Sometimes multiple antibiotics have to be tried before the patient feels better. Antibiotics may actually
help with the laboratory diagnosis. But patients need to be off antibiotics about 10 to 14 days before the
Western blot is repeated. This sounds like a contradiction. Antibiotics may help convert the test to
positive, but patients need to be off antibiotics when the specimen is drawn.
It is well documented in medical literature that the presence of antibiotics may cause false negative
borreliosis testing. Therefore, your system should be free of all antibiotics for an accurate blot result.
When the Lyme borrelia are alive, they are geniuses at avoiding the immune system. They may do things
like go inside your white blood cells, and come out enclosed by the cell membrane of your own white
blood cells! This may partly explain why antibodies against Borrelia burgdorferi are often not found when
patients are tested.
What may happen when patients are given 4 weeks of tetracycline (or other antibiotics) is that some of the
bacteria die. When Borrelia burgdorferi dies, it is less efficient at avoiding the immune system. That's
when antibodies may be formed against Borrelia burgdorferi, converting the negative or equivocal
Western blot to positive, in about 36% of cases.
If a borreliosis Western blot is going to be positive, it is usually the first one that is positive. The second
blot is the next most likely to be positive, and so on, until the fifth blot.
After that, the curve levels off for conversion to positive. This is based on research I presented in Bologna,
Italy in 1994. Some patients had borrelia-associated antibodies finally show on their tenth Western blot!
Two Western blots from a reliable lab usually gives the answer.
If a third test is needed, a Lyme Urine Antigen Test (LUAT) is done instead of a third Western blot.
Positive LUATs correspond very highly to patients getting better with antibiotics. False positive LUATs
have not been a problem in my practice. The LUAT finds the actual antigen (Borrelia burgdorferi itself),
so arguably it should be the test of choice, but the Western blot is rn6re widely accepted, even though it
looks for the antibodies against Borrelia burgdorferi.
The presence of antibodies are indirect evidence of an infection, not direct evidence like shown in the
LUAT. On the Western blot test result form, please note what is "considered positive" and "considered
equivocal." Equivocal is another way of saying suspicious or almost positive.
Below this are the ASTPHLD/CDC recommendations. The CDC stands for the Center for Disease
Control. I have been in attendance at the international borreliosis conferences when the CDC said their
recommendations are for disease surveillance, not day-to-day clinical medical practice. I am not in the
business of disease surveillance. My job is to try to help sick people.
The CDC recommendations do not include the 31 and 34 Kda bands of the blot test. These two bands
correspond to outer surface proteins A and B respectively (ospA and ospB). In the world of borreliosis,
these are two of the classic hallmark Lyme antibodies. But the CDC does not even have them in their
recommendations.
You may see why I and other borreliosis clinicians do not agree with using the CDC criteria in everyday
medical practice. Other bacteria besides Borrelia burgdorferi may produce the 45, 58, 66, and 73 kDa
bands. These bands may be produced by Borrelia burgdorferi, but are not nearly as specifically associated
with Lyme borreliosis as the starred bands. These starred bands are classic hallmark borrelia-associated
antigen-antibody complexes.
**An example of the CDC's criteria of a blot test, is if a patient has the band pattern of 41, 45, 58, 66, and
93, the CDC would call it positive. But if a patient has a 23-25, 31, 34, and 39 band pattern, they would
call it negative. This is despite the fact that this second pattern of antigen-antibody complex bands is
much more specifically associated with Borrelia burgdorferi than the first pattern.
As you can see, borreliosis is very controversial. It would be alarming if I was the only clinician who
thought that the CDC recommendations should not be used for day-to day medical practice. Many borrelia
clinicians do not use the CDC criteria. This is obvious by the fact that the IgX laboratory uses different
criteria for positive. Again, in my opinion and others', even one borrelia-associated antibody is significant,
if symptoms exist. The classic triad of symptoms for borreliosis is fatigue (tiredness, exhaustion),
musculoskeletal pain (joints, muscles, back, neck, headache), and cognitive problems (memory loss,
trouble concentrating, difficulty remembering what you read, depression, disorientation, getting lost).
But there are about 100 symptoms on the borreliosis questionnaire I use. Borreliosis may mimic or imitate
virtually any disease. Patients often tell me that other physicians they have seen use the CDC
recommendations. This is unfortunate, in my opinion, since these physicians are not in the business of
disease surveillance, like the CDC is.
But I am biased. After seeing patients with borreliosis since 1988, attending many conferences, talking
with experts, and doing research on borreliosis testing, there is absolutely no question in my mind that
physicians need to not blindly accept any recommendations.
One of my hopes is that doctors will someday realize that this controversy is a signal for them to search
for the truth. Why is there such conflict in this very "political" disease if there is not substance for
disagreement? Both IgG and IgM Western blots should be done for borreliosis.
With most infections, your immune system first forms IgM antibodies, then in about 2 to 4 weeks, you see
IgG antibodies. In some infections, IgG antibodies may be detectable for years. Because Borrelia
burgdorferi is a chronic persistent infection that may last for decades, you would think patients with
chronic symptoms would have positive IgG Western blots.
But actually, more IgM blots are positive in chronic borreliosis than IgG. Every time Borrelia burgdorferi
reproduces itself, it may stimulate the immune system to form new IgM antibodies. Some patients have
both IgG and IgM blots positive. But if either the IgG or IgM blot is positive, overall it is a positive result.
Response to antibiotics is the same if either is positive, or both. Some antibodies against the borrelia are
given more significance if they are IgG versus IgM, or vice versa.
Since this is a chronic persistent infection, this does not make a lot of sense to me. A newly formed
Borrelia burgdorferi should have the same antigen parts as the previous bacteria that produced it. But
anyway, from my clinical experience, these borrelia associated bands usually predict a clinical change in
symptoms with antibiotics, regardless of whether they are IgG or IgM. In regard to the outer surface
proteins, think of it like the skin of a human.
On the outer surface of the Lyme bacteria are various proteins. As they have been discovered, they have
been assigned letters, such as outer surface proteins A, B, and C. The following is a brief explanation of
the test results. Again, each band is an antigen complexed (bound together) with an antibody made by the
immune system, specifically for that antigen (part) of Borrelia burgdorferi.
18: An outer surface protein.
22: Possibly a variant of outer surface protein C.
23-25: Outer surface protein C (osp C).
28: An outer surface protein.
30: Possibly a variant of outer surface protein A.
31: Outer surface protein A (osp A). 34: Outer surface protein B (osp B).
37: Unknown, but it is in the medical literature that it is a borrelia-associated antibody. Other labs
consider it significant.
39: Unknown what this antigen is, but based on research at the National Institute of Health (NIH), other
Borrelia (such as Borrelia recurrentis that causes relapsing fever), do not even have the genetics to code
for the 39 kDa antigen, much less produce it. It is the most specific antibody for borreliosis of all.
41: Flagella or tail. This is how Borrelia burgdorferi moves around, by moving the flagella. Many bacteria
have flagella. This is the most common borreliosis antibody.
45: Heat shock protein. This helps the bacteria survive fever. The only bacteria in the world that does not
have heat shock proteins is Treponema pallidum, the cause of syphilis.
58: Heat shock protein.
66: Heat shock protein. This is the second most common borrelia antibody.
73: Heat shock protein.
83: This is the DNA or genetic material of Borrelia burgdorferi. It is the same thing as the 93, based upon
the medical literature. But laboratories vary in assigning significance to the 83 versus the 93.
93: The DNA or genetic material of Borrelia burgdorferi.
In my clinical experience, if a patient has symptoms suspicious for borreliosis, and has one or more of the
following bands, there is a very high probability the patient has borreliosis.
These bands are 18, 22, 23-25, 28, 30, 31, 34, 37, 39, 41, 83, and 93. This is true regardless of whether it
is IgG or IgM.. But again, there is (no universal agreement) on the significance of these bands. Betina
Wilska, M.D. from Germany is one of the world's experts on outer surface protein A (31 kDa).
At the international borreliosis conference in Vancouver, British Columbia, I asked her personally about
the 30 kDa band. She told me it was the same as the 31 kDa band (osp A). When you have the opportunity
to talk to borreliosis experts, this helps in assigning significance to findings, on an imperfect test. As a
medical doctor, I am stating all of this with no axe to grind, no professorship to protect, and no preset
opinions. Patients, personal research, and conferences have helped me interpret the borreliosis medical
literature in regard to testing. Nobody would like to have available a bullet-proof, 100% reliable Lyme
borreliosis test more than I would. But we must use what is currently available. I always welcome second
opinions.
Dr. C in Missouri



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