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Bb
Viestit: 1820
Liittynyt: Ma Tammi 26, 2009 23:13

VASTA-AINETESTI

Viesti Kirjoittaja Bb » Ti Helmi 10, 2009 00:24

Lähettäjä: iippo Lähetetty: 5.4.2004 11:52

Tietääkö kukaan tuottaako elimistö erilaiset vasta-aineet eri borrelia-lajeille, eli jos leikkisästi oletetaan, että esim. Elisa-testi olisi 100 % varma, niin onko kaikki alalajit testattava erikseen, vai paljastaako yksi testi kaikki borrelialajit?
_____________________________________________________________

Lähettäjä: Soijuv Lähetetty: 5.4.2004 13:14

Teitpä hankalan kysymyksen, enkä ole asiaan aiemmin törmännyt. Olen olettanut, että Elisa näyttää positiivista (silloin kun näyttäää), riippumatta alalajista. Näin ilmeisesti onkin - eli testi näyttää suurin piirtein positiivista riippumatta alalajista eli se reagoi spirokeettoihin ja joihinkin muihin samankaltaisiin mikrobeihin (tätä selitetään yleensä vääränä positiivisena vastauksena).

Näin on siis yleisesti ottaen, mutta esim. seuraavassa tutkimuksessa on todettu eri borrelia lajien aiheuttavan elimistössä erilaisia reaktiovoimakkuuksia. Tällöin on tietysti mahdollista että eri alalajit reagoisivat testiin herkemmin. En tunne asiaa paremmin - onko täällä ketään joka tuntisi asian paremmin?



LYMPHOCYTE PROLIFERATION ASSAY IN RESPONSE TO BORRELIA

BURGDORFERI S.L. IN EXPERIMENTALLY INFECTED C57bl/6 MICE

T. MALOVRH, J. JURCA, I. GRUNTAR, V. KOTNIK

Slov Vet Res 2002; 39 (3/4): 189-99

UDK 619:595.895.42:612.017:599.323.4

http://www.vf.uni-lj.si/veterina/zborni ... ovrh_s.pdf



Although there is a strong specific antibody-dependent and cellular immune response to the spirochaete in general, the illness cannot be successfully controlled in every patient .......
In this study we succeeded in showing that the different Borrelia strains elicited different levels of cell-mediated immune reactivity specific to the Borrelia strains used for immunisation. It is therefore clear that an unequivocal antigen-specific immune response may be expected only if the in vitro challenge antigen is of the same kind as that present in the spirochaete causing the infection.
___________________________________________________________


Lähettäjä: Sar_Ani Lähetetty: 8.4.2004 15:10

Hei ! WB:llä on monesti yritetty selvittää, mikä alalaji on kyseessä.

Laboratory Tests
Part 2: An Excerpt from the Lyme Disease Survival Manual
by Tom Grier

Western Blot

The Western Blot essentially makes a map of the different antibodies the immune system produces to the bacteria. The map separates the antibodies by the weight of their respective antigens and are reported in units called kilo daltons or kDa.

For example, a Western Blot may report bands at 22, 23, 25, 31, 34, 39, and 41 kDa. Each of these bands represents an antibody response to a specific protein found on the spirochete. The 41 band indicates an antibody to the flagella 41 kDa protein and is nonspecific. The 31 kDa band represents the OSPA protein and is specific for just a few species of Borrelia, as is the 34 band OSPB, and 23 kDa OSPC.

In 1994, the Association of State and Territorial Public Health Laboratory Directors, under a CDC grant, decided that there should be consistency between labs reporting Lyme disease Western Blots, and that a specific reporting criteria should be established. The consensus committe, chaired by Dr. Michael Osterholm, Ph.D., MN, set nationwide standards for Western Blot reporting. This sounds good, but one could argue they made a bad situation worse. Prior to the hearing, virtually every lab had accepted bands 22, 23, 25, 31, and 34 kDa as specific and significant, and reported them as positive for exposure to Borrelia burgdorferi. Not only are these bands specific for Borrelia species, but they represent all of the major outer surface proteins being used to develop the Lyme vaccines. The committee, without any clear reasoning, disqualified those bands as even being reportable.

After the consensus meeting, those bands were no longer acceptable. The result was that what had been a fair-to-good test for detecting Lyme disease had now become poor, arguably useless. Many scientists have questioned these new reporting criteria, and several wrote letters of protest to both the committee and to laboratory journals. Many labs stopped reporting the actual bands and instead, simply reported the test as positive or negative, thus preventing any further interpretations. (90)

How badly did the Lab Directors bootstrap this test? The following is an analysis of the new guidelines presented as an abstract and lecture at the 1995 Rheumatology Conference in Texas, chaired by Dr. Alan Steere, MD. (1995 Rheumatology Symposia Abstract #1254, Dr. Paul Fawcett, et al.)

This was a study designed to test the recently proposed changes to Western Blot interpretation by the Second National Conference on Serological Testing for Lyme Disease, sponsored by the CDC. The committee proposed limiting the bands that could be reported in a Western Blot for diagnosis of Lyme disease. Out of a possible 25 bands, 10 specific bands were selected as being reportable. An lgG Western Blot must have five or more of these bands: 18, 21,28, 30, 39, 41,,45, 58, 66 and 93 kDa. An lgM Western Blot must have two or more of the following three bands: 23, 39, 41.

Conspicuously absent are the most important bands, 22, 23, 25, 31, and 34, which include OSPA, OSP-B and OSP-C antigens - the three most widely accepted and recognized Bb antigens. These antigens were the antigens chosen for human vaccine trials. This abstract showed that, under the old criteria, all of 66 pediatric patients with a history of a tick bite and bull's-eye rash who were symptomatic were accepted as positive under the old Western Blot interpretation.

Under the newly proposed criteria, only 20 were now considered positive. (The number of false positives under both criteria was zero percent.) That means 46 children who were all symptomatic would probably be denied treatment! That's a success rate of only 31%.

* Note: A misconception about Western Blots is that they have as many false positives as false negatives. This is not true. False positives based on species specific bands are rare.

The conclusion of the researchers was: "the proposed Western Blot reporting criteria are grossly inadequate, because it excluded 69% of the infected children."

Elisa Test

The Enzyme-Linked Immunosorbant Serum Assay, is the simplest, least expensive, easiest to perform, and most common Lyme test ordered. It is a test based on detecting the antibodies that our bodies make in response to being exposed to Borrelia burgdorferi (Bb). It is a preferred test by laboratories, not because it is more accurate than other Lyme tests, but because it is automated. Many different patient samples can be performed by a single machine simultaneously. This allows for a faster turnover, less costs, and theoretically, standardized test results that are consistent from lab to lab.

We are told by manufacturers, health departments and clinics that the Lyme ELISA tests are good, useful tests, but in two blinded studies that tested laboratories for accuracy, they failed miserably. Lorie Bakken, MS/MPH, showed in her studies that there was not only inaccuracy and inconsistency between competing laboratories, but also between identical triple samples sent to the same lab. In other words, identical samples often resulted in different results! In the first study, forty-five labs correctly identified the samples only 55% of the time.

In the latest study by the College of American Pathologists, 516 labs were tested. The overall result was terrible! There were almost equal numbers of false positives as false negatives. Overall, the labs were 55% inaccurate. The labs could only give a correct result 45% of the time. You are actually better off to flip a coin!

The basis of the ELISA test is that it can be primed to be very specific for particular antibodies. This is done by taking a laboratory sample of the Lyme bacteria and breaking the sample down into fragments. These fragments, or antigens, are then embedded on the side of a reagent vessel like a test tube. Then the patient's serum is added, and any free (non-complexed) antibodies specific for the test strain will then bind to the antigens, which are linked to special enzymes that will change color when antibodies are present. The sample is continually diluted until the reaction no longer occurs and no color change can be detected. The sample is then reported as a dilution ratio, such as one part serum to 256 parts water, or 1:256.

The ELISA test sounds simple and straight forward, but it has a couple of major flaws. Borrelia species are some of the most polymorphic bacteria known to exist. In other words, most Borrelia species can significantly change its surface proteins enough during cell division as to evade our immune system, and may differ from laboratory strains enough to result in negative tests, even if antiBb antibodies are present! In Europe, this problem is intensified because they have recognized three species of Borrelia that cause Lyme disease, and so they have available three separate ELISA tests. The questions in America are: 1) Have we recognized all the strains and species of Borrelia that cause Lyme disease symptoms, and 2) are we incorporating them into our tests? The answer is no. Convenience and expedience has chosen that we don't prime our ELISA tests withwild strains, but use a laboratory strain.

When a lab reports that their ELISA test has had high specificity and high sensitivity, it is usually interpreted by doctors as being a more accurate test, but the doctors don't know what the lab is actually measuring. One of the hidden problems of serologic Lyme tests is the fact that the tests must be primed with a source of bacteria to create the reactions with the patient's antibodies. To do this, virtually all labs rely on a laboratory strain of Bb known as strain B-31.Taking purified antigens from strain B-31 and injecting them into mice, they then can extract a monoclonal antibody to each antigen, or a polyvalent antibody soup. This antibody is concentrated and purified, and then added to the ELISA test to test the efficacy and performance of the test. Unlike the wild strains, B-31 grows well in culture, and this makes it a perfect choice as a consistent and inexpensive source of Bb. But the affinity the mouse monoclonal antibody has to B-31 antigen is quite different from the affinity the patients' antibodies have to the same antigen. This means the test may register as negative because
the test cannot detect the slightly different antibody profile that a wild strain of Bb can produce. In other words, the labs are really comparing apples to oranges! This is why, when the American College of Pathologists used human sera to test the accuracy of 516 different laboratories ELISA tests nation wide, the overall accuracy was only 45%.

In the quest for specificity, most ELISA tests have become so specific that the test may fail to detect antibodies from related strains of Borrelia. This would include different genospecies that cause Lyme disease, as well as different Borrelia species that cause Tickborne Relapsing Fever. Would a cross reaction to the Borrelia species that cause Tick-borne Relapsing Fever be so bad?

The real Achilles' Heal of an ELISA Test is that it can only detect free antibody. It cannot detect any antibody that has become complexed with antigen.

The ELISA test depends on the active, free antibodies to attach to the free antigens that have been embedded on the walls of the test tube. If the antibodies in the serum being tested are already attached to antigens, then the enzyme reaction cannot take place. If we think of antibodies as sort of keys that fit into locks, and that on the surface of the bacteria are specific locks we now call antigens, you can see that once a key is inserted into a lock, the key is no longer available to open any other locks.

What makes this test so misleading is that many doctors accept high readings as an indication that the patient must really be sick. This logic is exactly backwards. If a patient is really infected with lots of bacteria, that means they have a lot of bacterial antigens floating around in the blood that are complexing free antibodies. So, as free antigen increases, free antibody decreases. Since the ELISA test detects only free antibody, a negative test might actually indicate a more serious infection. Many times, I have seen totally asymptotic patients with ELISA titers over 1000 be treated as though they were on death's doorstep simply because they had a high titer, while patients with borderline titers who are practically disabled are ignored, because a low titer is perceived as meaning less infected! These conclusions are erroneous and actually opposite to the truth, which is that a high titer means greater natural immunity.

This phenomena can actually be observed by using vaccines. If a patient has been vaccinated for a disease like tetanus, they will carry a high titer of free antibodies. If you try to measure those antibodies an hour after a booster shot is given, they will test negative. This is because the injected tetanus antigen complexes all available free antibody before the body can make more, so the measurable free antibody level drops.

The nature of all antibody is to seek out the proper antigen. The level of free antibody available is variable and often inadequate for the amount of antigen available. As antigen increases (i.e. The bacteria are dividing faster than the immune system can handle), free antibody drops.

What a high ELISA test may be a better indicator of is what level of immunity is the patient capable of mounting against this infection? A high titer is the same thing as saying the patient has a high natural immunity, and a low can mean that the patient may be overwhelmed by infection.

In one year-long study by Dr. Sam Donta, MD, done on chronic Lyme patients, the initial ELISA tests proved to be more than 66+% inaccurate (1996 LDF Conference lecture). Other researchers have also found the ELISA tests to be inaccurate. Using a 45-panel diagnostic testing protocol from the NIH for testing the efficacy of the ELISA and Western Blot, researchers found the accuracy of the Lyme ELISA varied from about 5075%, and were routinely inconsistent. The CDC's ELISA test did no better on average than any other ELISA. It is the CDC ELISA test which is used for surveillance of emerging Lyme disease in the United States, yet the test was correct only about two out every three tests. Too often, a single negative ELISA test can prevent a sick patient from getting treatment, even despite having serious symptoms!

In my opinion, the ELISA test is worthless as a diagnostic tool in Lyme disease. It is inconsistent and inaccurate, and should be discontinued as a tool to diagnose Lyme. If the NIH and CDC truly believe, as they've stated, that the diagnosis of Lyme disease is to be made on the basis of symptoms, then these tests should be temporarily banned until each manufacturer can prove efficacy using human serum.

http://www.geocities.com/HotSprings/Oas ... n-blot.txt

And the bands played on - Western blot serological test for Lyme disease
************************************************************************
as of 4 October 1999
Table of Contents
Terminology.
Borrelia burgdorferi bands mentioned in medical literature (MEDLINE).
Named bands - the gene names.
Bands specific for Borrelia burgdorferi.
Cross-reacting bands.
Bands found, or tested for, in IgM analysis in USA or Mexico.
Bands found, or tested for, in IgG analysis in USA or Mexico.
Other bands found in IgM/IgG in other countries.
The "CDC recommended" bands for Western blot testing - CDC/MMWR 1995.
For more information about the Western blot test and Lyme disease.
-----
Terminology:
Bb Borrelia burgdorferi (the Lyme disease bacteria)
Bdr Borrelia direct repeat
Bmp Bacterial membrane protein
Dbp decorin binding protein
Fla flagellin
Fn-B fibronectin-binding protein
GroEL a chaperonin 60 heat-shock protein isolated from Escherichia coli
Hsp heat shock protein
HSP Heat Stress Protein
kDa kilodaltons (a measurement of size)
Mab monoclonal antibodies
MgtE magnesium transporter protein
Oms outer membrane-spanning
Osp outer surface protein
P protein
p protein
PBP penicillin-binding protein
PMID PubMed ID (identification system for citations)
Tbp transferrin-binding protein
-----
Borrelia burgdorferi bands mentioned in medical literature (MEDLINE):
Note: Bands preceded with an asterisk are the 11 Western blot bands for
the ASTPHLD, CDC, FDA, NIH, CSTE, NCCLS 1994 conference recommendation
("CDC recommendation") for the serologic diagnosis of Lyme disease -
see 1995 CDC MMWR link below.
5-kDa
7.5-kDa
11-kDa
13-kDa surface protein - sensu stricto, afzelii
14-kDa internal flagellin fragment [specific for Bb]
15 kDa polypeptide [also for syphilis]
16-kDa
17-kDa Osp 17 [B. afzelii]
*18-kDa p18 flagellin fragment
19-kDa immunogenic integral membrane lipoproteins
cross-reactive with other spirochetes/bacteria
Characterization of antigenic determinants of Bb shared by other
bacteria. http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
19-kDa decorin-binding protein
20-kDa decorin-binding protein
20.7-kDa
*21-kDa OspC [specific for Bb]
22-kDa [specific for Bb or cross-reactive depending on what one reads]
immunogenic integral membrane lipoproteins
[cross-reactive with other spirochetes/bacteria]
Characterization of antigenic determinants of Bb shared by other
bacteria. http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
22-kDa OspC [specific for Bb]
22-25kDa OspC
23-kDa OspC
*24-kDa OspC
25-kDa OspC [specific for Bb]
26-kDa
25-kDa
27-kDa Osp, Hsp
(Europe burgdorferi strain B29, but not American strain B31)
*28-kDa OspD, Oms28 [specific for Bb]
29-kDa
30-32-kDa OspA
*30-kDa OspA substrate binding protein
31-kDa OspA [specific for Bb]
32-kDa OspA
33-kDa outer membrane
34-kDa OspB [specific for Bb]
34-36-kDa OspB
35-kDa OspB [specific for Bb]
35.5-kDa
36-37-kDa
37-kDa P37, flaA gene product, [specific for Bb]
38.0-kDa
*39-kDa BmpA [specific for Bb]
40-kDa
*41-kDa Fla
42-kDa
43-kDa
44-kDa
*45-kDa [appeared in IgM in control group in 1998 study done in Poland]
MEDLINE - 9972057 - "...whereas in control group only antibodies
against 45 kDa and 58 kDa were present." http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
[appears for HGE]
MEDLINE - 9620365 - "...confirmed the importance of the 42- to
45-kDa antigens as early, persistent, and specific markers of HGE
infection." http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
46-kDa
47-kDa P47 fibronectin-binding protein [specific for Bb]
48-kDa
49-kDa
50-kDa [specific for Bb]
51 kDa MgtE
52-kDa Fn-BA
54-kDa [other Borrelia]
55-kDa
56-kDa
57-kDa PBP
*58-kDa (not GroEL)
59-kDa [a genetically engineeried fragment of the 83-kDa protein]
60-kDa Hsp [all Borrelia]
62-kDa Hsp60
63.7-kDa
64-kDa (P64) [cross-reacts to human axonal proteins]
65-kDa
*66-kDa P66 Oms66 Hsp outer/integral membrane protein
67-kDa
68-kDa
70-kDa Hsp
71-kDa
72-kDa Hsp [cross-reactive with other spirochetes]
[cross-reactive with other spirochetes/bacteria]
Characterization of antigenic determinants of Bb shared by other
bacteria. http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
73-kDa
75-kDa
77-kDa a genetically engineered recombinant hybred
Use of a hybrid protein consisting of the variable region of the
Borrelia burgdorferi flagellin and part of the 83-kDa protein as
antigen for serodiagnosis of Lyme disease. http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
78-kDa OspA
79.8-kDa
80-kDa
83-kDa p83 high molecular mass protein [specific for Bb]
84-kDa [B. garinii]
88-kDa
92-kDa
*93-kDa an immunodominant protoplasmic cylinder antigen, associated with
the flagellum [specific for Bb]
94-kDa PBP [specific for Bb]
95-kDa
97-kDa associated with flagella
100-kDa P100
110-kDa
200-kDa a fusion protein, a hybrid protein
-----
Named bands - the gene names:
BmpA, BmpB, BmpC "the 39 kDa Bmp protein family", PMID: 8978084
BmpD, PMID: 8606088
FlaA osp of the periplasmic flagella, 37-kDa, PMID: 9986810;
38-kDa, PMID: 9573194, PMID: 8990312
FlaB 41-kDa PMID: 9573194
FlgE protein 40-kDa?, PMID: 8548542
ospAB 28-34-kDa, PMID: 9596714
OspA 31-kDa, PMID: 10030131
OspB 34-kDa, PMID: 10030131
OspC 21-25-kDa, 22-25-kDa - PMID: 8825913,
OspD 28-kDa, PMID: 8825913
OspE 19.2-kDa [calculated], PMID: 8262642
OspF 26.1-kDa [calculated], PMID: 8262642
----
Bands specific for Borrelia burgdorferi:
14-kDa ?
21-kDa
22-kDa OspC
25-kDa OspC
28-kDa OspD
31-kDa OspA
34-kDa OspB
35-kDa
37-kDa
39-kDa
47-kDa
50-kDa
83-kDa
93-kDa
94-kDa
Notes:
14-kDa ? PMID: 9920119, PMID: 1556546 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
B. burgdorferi: 22 kDa, 31 kDa, 34 kDa, 39 kDa, 83 kDa - PMID: 9440203 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
p35 and p37 are Borrelia burgdorferi genes - PMID: 9175831 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
at least one is an apparently specific band (25, 28, 39, 47, 50, or
93 kDa). - PMID: 7791177 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
antibodies against the 94 kDa, 31 kDa and 21 kDa proteins are largely
species-specific. - PMID: 8223404 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
-----
Cross-reacting bands:
19-kDa
22-kDa ??
20-kDa
30-kDa
33-kDa
34-kDa ??
36-kDa
40-kDa
41-kDa
60-kDa
66-kDa
72-kDa
Notes:
Search terms:
cross-react*
cross-antigenicity
crossreact
Immunoblot analysis indicated the presence of cross-reacting antibodies
directed to B. burgdorferi antigens with apparent molecular weights of
60, 41, 40, 36, 30 and 20 kDa. - PMID: 1385332 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
P66, P60, P41 which are dominant immunogens of all types of borrelias
PMID: 9162453 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
19, 22[??], 72 - PMID: 1372635 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
60-75 kDa range, p40, p33 and two proteins in the range of 20 kDa. -
PMID: 1597198 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
Whereas the 60 kDa, 41 kDa, and 34 kDa[??] constituents reveal a
marked cross-antigenicity with other spirochetes and even more distantly
related bacteria,...PMID: 8223404 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
-----
Bands found, or tested for, in IgM analysis in USA or Mexico.
18-kDa
21-kDa
23-kDa
24-kDa
25-kDa
28-kDa
37-kDa
39-kDa
41-kDa
45-kDa
55-kDa
58-kDa
60-kDa
66-kDa
93-kDa
Notes:
37 - PMID: 9986810 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
41 kDa and 58 kDa - PMID: 9972057 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
24 kDa (OspC), 41 kDa, and 37 kDa - PMID: 8748261 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
39, 58, 60, 66, or 93 kDa - PMID: 8748261 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
55-kDa - PMID: 7642278 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
Recommended criteria for the immunoglobulin M (IgM) immunoblot are the
recognition of two of three proteins (24, 39, and 41 kDa). PMID: 7714202 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
25-kDa antigens - PMID: 8308100 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
23-kDa - PMID: 8225587 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
at least 2 of the 8 most common IgM bands in early disease
(18, 21, 28, 37, 41, 45, 58, and 93 kDa) - PMID: 8380611 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
-----
Bands found, or tested for, in IgG analysis in USA or Mexico.
18-kDa
20-kDa
21-kDa
22-kDa
23-kDa
24-kDa
28-kDa
29-kDa
30-kDa
31-kDa
34-kDa
35-kDa
39-kDa
41-kDa
45-kDa
55-kDa
58-kDa
88-kDa
62-kDa
66-kDa
93-kDa
Notes:
A serum sample was considered positive by WB [IgG] if at least three of
the following protein bands were recognized: 18, 24, 28, 29, 31, 34, 39,
41, 45, 58, 62, 66, and 93 kDa. PMID: 10071428 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
93, 39, 34 or 23 kDa IgG bands - PMID: 9580180 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
55-kDa - PMID: 7642278 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
The recommended criteria for a positive IgG immunoblot are the
recognition of two of five proteins (20, 24 [> 19 intensity units], 35,
39, and 88 kDa). Alternatively, if band intensity cannot be measured,
the 22-kDa protein can be substituted for the 24-kDa protein with only
a small decrease in sensitivity. PMID: 7714202 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
at least 5 of the 10 most frequent IgG bands after the first weeks of
infection (18, 21, 28, 30, 39, 41, 45, 58, 66, and 93 kDa)-PMID: 8380611 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
66 kDa and 31/34 kDa - PMID: 3819479 http://www.ncbi.nlm.nih.gov/htbin-post/ ... b=m&Dopt=b
-----
Other bands found in IgM/IgG in other countries:
Europe/Germany - PMID: 9163458
The following interpretation criteria resulting in specificities of
greater than 96% were recommended: for IgG WB, at least one band of
p83/100, p58, p56, OspC, p21, and p17a for PKa2; at least two
bands of p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for
PKo; and at least one band of p83/100, p39, OspC, p21, and p17b
for PBi; for IgM WB, at least one band of p39, OspC, and p17a or
a strong p41 band for PKa2; at least one band of p39, OspC, and
p17 or a strong p41 band for PKo; and at least one band of p39 and
OspC or a strong p41 band for PBi.
IgG titers to P35 and P37 Mice/Northern blot - PMID: 9175831
Italy - PMID: 8809552
The overall reactivity of the
three Borrelia strains in IgG immunoblots indicated that ten
protein bands were significant, with a different prevalence of some
of them in the two groups of patient sera: bands at 60-58, 30-33,
36-37 and 28-27 kDa were markers for neuroborreliosis sera;
proteins at 100-83, 72-70 and 18-17 kDa behaved like markers for
Lyme arthritis. The IgM Immunoblots revealed significant bands
at 100-83, 72-70, 51, 24- 21 and 18-17 kDa only with
neuroborreliosis sera.
Russia - PMID: 7791165
IgG reactivity with > or = 5 spirochetal proteins, particularly the
37, 39, 41, 60, and 93 kDa antigens
Germany - PMID: 8167425
IgG - 95 and 19-17 kDa
France - PMID: 8405312
[IgG] Reactions with specific protein bands (94, 73, 30 and 21 KDa)
-----
The "CDC recommended" bands for Western blot testing - CDC/MMWR 1995.
The ASTPHLD, CDC, FDA, NIH, CSTE, NCCLS 1994 conference recommendation
("CDC recommendation") for the serologic diagnosis of Lyme disease -
CDC/MMWR 1995:
ASTPHLD - Association of State and Territorial Public Health Laboratory
Directors
CDC - Centers for Disease Control and Prevention
FDA - Food and Drug Administration
NIH - National Institutes of Health
CSTE - Council of State and Territorial Epidemiologists
NCCLS - National Committee for Clinical Laboratory Standards
According to CDC's Morbidity and Mortality Weekly Report (MMWR) 1995;
44 (31):590-591, an IgM immunoblot is considered positive if two of the
following three bands are present:
24 kDa (OspC)*
39 kDa (BmpA)
41 kDa (Fla)
An IgG immunoblot is considered positive if five of the following 10
bands are present:
18 kDa
21 kDa (OspC)*
28 kDa
30 kDa
39 kDa (BmpA)
41 kDa (Fla)
45 kDa
58 kDa (not GroEL)
66 kDa
93 kDa
* The apparent molecular mass of OspC is dependent on the strain of
B. burgdorferi being tested. The 24 kDa and 21 kDa proteins referred
to are the same.
Recommendations for Test Performance and Interpretation from the Second
National Conference on Serologic Diagnosis of Lyme Disease,
CDC MMWR, August 11, 1995 / 44(31);590-591 http://www.cdc.gov/epo/mmwr/preview/mmw ... 038469.htm
or http://wonder.cdc.gov/wonder/prevguid/m ... entire.htm
-----
For more information about the Western blot test and Lyme disease, see:
The National Lyme Disease Network LymeNet Newsletter
Volume 4 - Number 13 - 9/23/96
SPECIAL ISSUE - Understanding the Western Blot http://www2.lymenet.org/domino/nl.nsf/UID/4-13
Explanation of the Lyme Disease Western Blot by Carl Brenner http://www.lymealliance.org/Medical/Med ... /lab4.html
Laboratory Tests by Tom Grier - Part 2 - Western Blot and ELISA http://www.lymealliance.org/Medical/Med ... ed12a.html
IGeneX, Inc. - Lyme Disease Western Blot http://www.igenex.com/lymeset2.htm
MRL Diagnostics' Lyme Disease B. burgdorferi Genogroup 1
Western Blot IgG test http://www.mrldiagnostics.com/insert/wb0400g.htm
Immuno-Biological Laboratories (IBL) - Hamburg [Germany] -
Borrelia burgdorferi/Lyme IgG Western Blot http://www.ibl-hamburg.com/prod/re_97228.htm
The western immunoblot for Lyme disease: determination of sensitivity,
specificity, and interpretive criteria with use of commercially
available performance panels.
Tilton RC, Sand MN, Manak M
BBI Clinical Laboratories, Inc., New Britain, Connecticut 06053, USA.
Clin Infect Dis 1997 Jul;25 Suppl 1:S31-S34 http://www.x-l.net/Lyme/BBI_TEST.htm
SIMULTANEOUS ELISA AND WESTERN BLOT TESTING IN EVALUATION OF PATIENTS
FOR SUSPECTED LYME DISEASE
Janice M. Kochevar, FNP-C, Kenneth B. Liegner, M.D.
LDF 10th Annual International Scientific Conference on Lyme Borreliosis
& Other Tick-borne Disorders: April 28-30, 1997 http://www.x-l.net/Lyme/elisawb497.htm
MEDLINE - Western Immunoblotting/immunoblots/blottting/blots [in Title]
AND Lyme Disease - 31 on 20 Aug 99 http://igm-02.nlm.nih.gov/cgi-bin/IGM_r ... lot+OR+Tit le+Word=Western+immunoblots+OR+Title+Word=Western+immunoblot+AND+Subject=lyme+OR+Subject=burgdorferi+OR+Subject=Erythema+Chronicum+Migrans+OR+Subject=borreli*+ixodes+OR+Subject=Ery thema+Migrans+NOT+glossitis+OR+Subject=garinii&datafile=MEDLINE
-----
And, for more information on Lyme disease, see:
Lots Of Links On Lyme Disease http://www.geocities.com/HotSprings/Oas ... links.html
_____________________________________________________________


Lähettäjä: iippo Lähetetty: 8.4.2004 15:55

Moi
kiitos vastauksesta. Muistan lukeneeni tämän saman jutun jostakin kun tuo "You are actually better off to flip a coin!"-vertaus jäi niin hyvin mieleen, mutten tuolloin oikein sisäistänyt asiaa.

Pitää ottaa näitä tärkeitä asiapapereita mukaan kun ensi viikolla menen lääkärin pakeille.

Iloista pääsiäistä kaikille!
_____________________________________________________________


Lähettäjä: jukka61 Lähetetty: 11.4.2004 19:14

Moi!

Kuinkakohan monelle on Western Blot tehty ja kuinka hyvin se on osattu tulkita?

Minulle on tehty 3-kertaa ja aika tuttuja numeroita tuon artikkelin perusteella esiintyi testeissä.

Siitä huolimatta, että noi Western blot ovat olleet "positiivisia", samoin, kun joka ikinen verikoe (enemmän kuin 15kpl), selkäytimestä löytyi tulehdusta, jne. ja liikun ja vietän (liikuin ja vietin) yli 100 päivää punkkialueella Inkoossa ja Tammisaaressa, jossa punkkeja on luonnollisesti ollut kiinni, niin siitäkin huolimatta diagnoosin kanssa on vitkuteltu, asia on kyseenalaistettu, pitkään kuljin reumadiagnoosilla, itse asiassa ensi viikolla tutkitaan taas erilaisilla kuvauksilla tulehdusten ja reuman mahdollisuutta.

Hyvä niin, mutta jostain syystä Borren kroonistumista ja uusiutumista ei meinata millään uskoa ja hyväksyä, ikään kuin kyseessä olisi pyhä ja periaatteellinen asia.

Huvittavaksi asian tekee viekä se, että en itse koe tulehduksia ja kipuja (nivelissä, jne.) ollenkaan niin kiusalliseksi, kuin erilaisia neuro- ja "sisuskalu"oireita, jotka pahimmillaan lamaavat täysin. Tämä asia on hemmetin vaikea selittää lääkäreille, noille neuro-oireille, kun ei ole oikein kunnon tutkimusmenetelmiä.

Vastaa Viestiin