TerryK
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I just got some test results for my lyme peptide test and am trying to understand them. I'll be discussing the results with my doctor in a few weeks but I want to understand the results enough to know what questions I should be asking.
See below for more information on the test itself. It is a combination of 2 types of ELISA and the Western Blot. If any of you are familiar with this test and can answer questions or point me to more information, I'd really appreciate the help.
Reference ranges are all 0-2 meaning that anything above a 2 is abnormal. I've only included the positive results.
From what I can tell in reading some of the text in my test results, this *may* indicate that I have babesia. By reading the text, I'm not sure if one must have both IgG and IgM for that or if only IgG alone would qualify.
I had no Borellia Garinii or Afzelii positives on this test although I've had quite a few borellia specific bands show up on on my IgeneX and MDL Western Blots. None have been CDC positive and one Western Blot was IgeneX IgG positive.
"Lyme Antibodies (IgG/IgM) Unrelated Peptide - An unrelated peptide is tested to give an indication of the propensity of the humoral immune system to be prone to molecular mimicry/cross reactivity (i.e. the propensity to be unable to differentiate from among antigens that are similar in structure and/or composition)."
According to the same link: The babesia Peptide & Erlichia Peptide are antigenically similar to Borrelia burgdorferi and may cause cross reactive antibody production to Borrelia burgdorferi peptides.
Here are my questions: 1. Are there any other known infections that cause similar antigens and can therefore cross react? If so, what are they?
2. What, are the implications that I tested above the range for "unknown spirochete"? a. Does this imply that my immune system may be causing me to erroneously test positive on some of the Western Blot bands that are specific to Bb?
For those who are interested, here is the text about the test itself (split into paragraphs to make it easier to read):
"...Therefore, it is critical to combine clinical symptomatology with the most sensitive technique available to diagnose Lyme Disease. In search of such a sensitive method, we applied the commercially available ELISA, Western Blot and the newly-developed peptide-based ELISA in our laboratory.
Since the chronic nature of Lyme disease and antigenic diversity of the spirochetes suggest that antigenic variation plays an important role in immune invasion, we selected peptides from different components of Borellia during different cycles, including peptides from outer surface protein leukocyte function associated antigens, immunodominant antigens, variable major proteins and peptides from decorin-binding proteins of Borellia species (B. sensu stricto, B. afzelii, B. garinii).
Furthermore, in the same ELISA assay, we measured antibodies against specific peptides from Babesia and Erlichia in order to exclude cross-reactive antibodies."
Thanks for any input.
Terry
Posts: 6286 | From Oregon | Registered: Jan 2006
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Was the Immunoserology of Lyme Disease the test you had done? My PC could not load this file. I called and the lab sent me a brochure. Did not give a sample test or any info that I can compare to your results.
Hubby plans to do this test in mid June -- has had to reschedule appointment with PCP 2 times or would have already had the test.
Would also suggest you call the lab. Let us know if you find out more.
Bea Seibert
Posts: 7306 | From Martinsville,VA,USA | Registered: Oct 2004
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bettyg
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see my 4-20-06 LLMD summary & lab results where I updated even more today.
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Haven't heard of this test before. On the face of it, seems like anything that combines several tests ought to be an improvement, but I just don't know enough to evaluate it. Guess it is time to dig into the matter further, or hope someone comes along, like clinlabscientist to explain why this might work better or is not any more accurate. Doesn't this still depend on antibodies being produced? As we know, some people are seronegative on antibody tests in chronic cases, but then turn positive with treatment when their immune systems come back to life, so to speak.
Both the ELISA and WB and C6 peptide tests are measuring antibodies. What is that LFA antigen doing in the list?
Posts: 8430 | From Not available | Registered: Oct 2000
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treepatrol
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C6 Peptide Assay
C6 is a synthetic peptide {C6 Peptide} derived from the VISE protein which appears in early and late Lyme Disease. The assay looks for the presence of antibodies against this synthetic peptide. While not as sensitive as the IGeneX Western Blots, it has no demonstrated cross reactivity in patients who may have received the LymeRix� vaccine.
Starting October 2003, IGeneX is offering the FDA approved Immunetics C6 Assay. The C6 B. burgdorferi {Lyme} ELISA Kit assay is intended for use in the presumptive detection of lgG and lgM antibodies to B. burgdorferi in human serum. The assay is approved for use on samples from patients with clinical history, signs or symptoms consistent with B. burgdorferi infection, including individuals who have received the licensed recombinant OspA Lyme disease vaccine (LymeRix�).
A set of 27 patient sera suspected of having Lyme Disease were tested by the C6 assay and Lyme Western blot IgG and IgM assays. The Lyme IgG and IgM Western Blots, were scored positive or negative based on the IGeneX criteria {WB --IGeneX} and by the CDC criteria.
14 {52%} of the 27 patient sera were positive by the C6 assay. All 14 were positive by the Western Blot assay, by both IGeneX Criteria (WB--IGeneX) and CDC criteria {WB--CDC}. In addition, 5 more were positive by the IGeneX Western Blot criteria. Of these, 4 were also positive by the CDC criteria.
As shown in the graph above, based on the study performed at IGeneX, the assay sensitivity of the C6 assay was between 70 and 74%, as compared to Western Blots. Thus, we continue to recommend that all patients' sera be tested by a Western Blot method.
Positive Western Blot results provide evidence for exposure to or infection with B. burgdorferi. The diagnosis of Lyme disease must be made based on history, signs {such as erythema migrans}, symptoms, and other laboratory data, in addition to the presence of antibodies to B. burgdorferi. Negative results should not be used to exclude Lyme disease.
The only reason for the c6peptide test in my opinion should be if you had lymerix vacine.
AND Ability to detect antibodies to all US and European strains of B. burgdorferi, because this protein is common to all presently known strains of the bacteria.
Patients with autoimmune diseases {e.g. non-Lyme arthritis, lupus erythematosis}, other spirochetal diseases {syphilis, relapsing fever}, neurologic conditions, and infectious diseases. Because the C6 LPE is highly specific, people with autoimmune diseases do not yield false positive results as they often do with current Lyme tests.
-------------------- Do unto others as you would have them do unto you. Remember Iam not a Doctor Just someone struggling like you with Tick Borne Diseases.
SForsgren
Frequent Contributor (1K+ posts)
Member # 7686
posted
Some thoughts:
1) If you have symptoms, these tests seem to support chronic infection in my opinion. 2) They only test for Babesia Microti. So I would say that is a positive test and there could also be other Babesia involved that they do not test for. 3) Immunosciences test is NOT a good test for coinfections. I would get coinfection tests done elsewhere. They only test for one stain of each, thus potentially missing some of the strains that are also common, especially the Ehrlichias. I personally find the test misleading in that they call it a test for "Coinfections" when in reality it is really not. They are looking for cross-reactivity with Lyme, but not really attempting to do a true coinfection test. 4) IGeneX and other labs may not find Afzelii and Garinii. The jury is still out on whether or not they would find them or not. They are only looking for Borrelia Burdgorferi. That is one area where I do like the Immunosciences test. 5) I would think it unlikely to have other infections that cross-react. They test for those in this test. Trepenoma for example. 6) I am not sure what "unknown spirochete" means. Call Immunosciences or email them and ask to talk to Dr. Vojdani if you want to get more information. I would love to hear what that means as well.
Be well, Scott
-------------------- Be well, Scott Posts: 4617 | From San Jose, CA | Registered: Jul 2005
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treepatrol
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A little more reading: The C6 Lyme Peptide ELISA A new Lyme disease test is called the C6 Lyme Peptide ELISA.
While it is also an antibody test, which assumes the antibody system is working well, is has unique aspects.
Lyme has the ability to have many forms and strains. It has ability to hide from the immune system. This test detects antibodies to a newly discovered consistent protein on every known strain of the Lyme disease bacteria.
According to the manufacturer and some researchers, Lyme is able to evade immune system response by many means, including varying its coat seen by our immune system. Much like wearing a different coat in the winter to escape detection after robbing a bank�this allows it "variation."
How often does this change in the Lyme coat happen? One study notes Lyme varies its coat or other key proteins on certain days. The clock for coat changes starts after infection in mammals after initial infection on days 4,7,14,21,28, and 7 and 12 months post infection of mammalian hosts. {Zhang, Norris�abstract below}
According to the maker, scientists have found a part of the Lyme "coat" that does not appear to change. Specifically, "6 surface proteins" on Lyme remain constant. Scientists call these "invariable regions" {IR}, which are termed IR1-6. One IR, IR6 is found in all Lyme strains. The new test is abbreviated, C6 LPE, and detects antibodies to the IR6 in infected patients.
The maker of the test reports: "diagnosis of Lyme disease has been hampered by tests that have not been standardized, are not reproducible, and are neither sensitive nor specific for this often difficult to diagnose tick-borne disease. The recent Lyme disease vaccine, which has been given to thousands of people over the past year, makes current tests for Lyme disease virtually useless. The C6 LPE has several features that ensure its accuracy and reproducibility in a wide range of patients."
BBI Clinical Laboratories, the maker of this lab test, says this new test is useful for:
Patients that have received the Lyme disease vaccine. The ability to detect antibodies to all US and European strains.
Patients with autoimmune diseases {e.g. non-Lyme arthritis, lupus erythematosis}, other spirochete diseases (syphilis, relapsing fever), neurologic conditions, and infectious diseases. Because the C6 LPE is highly specific, people with autoimmune diseases do not yield false positive results as they often do with current Lyme tests. Since IR6 antibodies begin to be produced in early infection and are produced throughout the course of infection, the C6LPE is more sensitive for diagnosing all stages of Lyme disease, including those patients with late stage Lyme disease.
Critique and Concerns by Some Clinicians. Some researchers feel this is a useful contribution to the lab options for testing for Lyme. It is moving through the FDA approval process reasonably as far as I can tell.
Currently, the junk way of testing for Lyme is to check a general titer and the check for the Western Blot "bands." This is called the "two-tiered" approach. While this is a poor system, it is at least occasionally useful, and can allow doctors to think through the values. Meaning, if the clinical picture makes Lyme likely and the tests are marginal, some doctors would feel Lyme is likely positive and treat.
If it replaces the other tests, we are back to the simplistic basic approach to Lyme. "Oh, one test will answer all our questions."
One scientist he is not so sure it will pick up late stage patients. I do not know if this is correct.
Other's doubt that the antibody levels will remain constantly high and measurable for months and years.
If it is done, especially as the only test, if it comes back negative, even if the patient has Lyme rashes all over them, it will be hard to get any insurance company to pay for treatment. They will merely say, "Hey, the C6 Elisa test is negative so we reject your care."
Kinetics and in vivo induction of genetic variation of vlsE in Borrelia burgdorferi.
Zhang JR, Norris SJ.
Department of Pathology and Laboratory Medicine and Department of Microbiology and Molecular Genetics, University of Texas Medical School at Houston, Houston, Texas 77030, USA.
The Lyme disease agent, Borrelia burgdorferi, is able to persistently infect humans and animals for months or years in the presence of an active immune response. It is not known how the organisms survive immune attack in the mammalian host. vlsE, a gene localized near one end of linear plasmid lp28-1 and encoding a surface-exposed lipoprotein in B. burgdorferi B31, was shown recently to undergo extensive genetic and antigenic variation within 28 days of initial infection in C3H/HeN mice.
In this study, we examined the kinetics of vlsE sequence variation in C3H/HeN mice at 4, 7, 14, 21, and 28 days and at 7 and 12 months postinfection. Sequence changes were detected by PCR amplification and sequence analysis as early as 4 days postinfection and accumulated progressively in both C3H/HeN and CB-17 severe combined immunodeficient {SCID} mice throughout the course of infection. The sequence changes were consistent with sequential recombination of segments from multiple silent vls cassette sites into the vlsE expression site.
No vlsE sequence changes were detected in organisms cultured in vitro for up to 84 days. These results indicate that vlsE recombination is induced by a factor{s} present in the mammalian host, independent of adaptive immune responses. The possible inducing conditions appear to be present in various tissue sites because isolates from multiple tissues showed similar degrees of sequence variation. The rate of accumulation of predicted amino acid changes was higher in the immunologically intact C3H/HeN mice than in SCID mice, a finding consistent with immune selection of VlsE variants.
Genetic variation of the Borrelia burgdorferi gene vlsE involves cassette-specific, segmental gene conversion.
Zhang JR, Norris SJ.
Department of Pathology and Laboratory Medicine and Department of Microbiology and Molecular Genetics, University of Texas Medical School at Houston, Houston, Texas 77030, USA.
The Lyme disease spirochete Borrelia burgdorferi possesses 15 silent vls cassettes and a vls expression site {vlsE} encoding a surface-exposed lipoprotein. Segments of the silent vls cassettes have been shown to recombine with the vlsE cassette region in the mammalian host, resulting in combinatorial antigenic variation. Despite promiscuous recombination within the vlsE cassette region, the 5' and 3' coding sequences of vlsE that flank the cassette region are not subject to sequence variation during these recombination events.
The segments of the silent vls cassettes recombine in the vlsE cassette region through a unidirectional process such that the sequence and organization of the silent vls loci are not affected. As a result of recombination, the previously expressed segments are replaced by incoming segments and apparently degraded. These results provide evidence for a gene conversion mechanism in VlsE antigenic variation.
Antigenic variation in Lyme disease borreliae by promiscuous recombination of VMP-like sequence cassettes.
Zhang JR, Hardham JM, Barbour AG, Norris SJ.
Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston 77030, USA.
We have identified and characterized an elaborate genetic system in the Lyme disease spirochete Borrelia burgdorferi that promotes extensive antigenic variation of a surface-exposed lipoprotein, VlsE. A 28 kb linear plasmid of B. burgdorferi B31 {lp28-1} was found to contain a vmp-like sequence {vls} locus that closely resembles the variable major protein (vmp) system for antigenic variation of relapsing fever organisms.
Portions of several of the 15 nonexpressed {silent} vls cassette sequences located upstream of vlsE recombined into the central vlsE cassette region during infection of C3H/HeN mice, resulting in antigenic variation of the expressed lipoprotein. This combinatorial variation could potentially produce millions of antigenic variants in the mammalian host.
PMID: 9108482 {PubMed - indexed for MEDLINE}
-------------------- Do unto others as you would have them do unto you. Remember Iam not a Doctor Just someone struggling like you with Tick Borne Diseases.
Don't think the Immunosciences test is the same as the C6 Peptide test.
As I said, my computer is too slow(dialup) to load the info that explains the test. Click on the link that says Immunoserology of Lyme to read more about this specific test.
-------------------- Do unto others as you would have them do unto you. Remember Iam not a Doctor Just someone struggling like you with Tick Borne Diseases.
SForsgren
Frequent Contributor (1K+ posts)
Member # 7686
posted
Just remember that the Immunosciences test is NOT a reliable indication of lack of coinfection. They do not test for all of the testable strains.
-------------------- Be well, Scott Posts: 4617 | From San Jose, CA | Registered: Jul 2005
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TerryK
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Thank you all for your responses. I called the lab as many of you suggested and they will not discuss even general questions. After much discussion, the receptionist checked with the lab techs and they told her to tell me that "unknown spirochete" is a bacteria. LOL According to the person who answers the phone, Dr. Vojdani does not talk to patients but will talk to me if I have my doctor on the phone with me. This is the standard response that I have gotten from many labs. After much pushing, the receptionist said she would have a lab person call me today or tomorrow. I won't hold my breath.
tj - I had been to their website but I was looking for lyme peptide A (which is what my doctor ordered) rather than the correct test of immunoserology of lyme. Welcome back to Portland!
Bea - Thank you so much for the link. Very helpful. I was able to download the information. I will send it in an attachment to you if you like. I can also send it by copying the contents of the pdf and pasting it into an e-mail but the formatting makes it hard to read when done that way. Yes, the test I had is called the immunoserology of Lyme-A.
bettyg - Is there something in your test results that relate to this test? I looked through your results but saw no mention of this test.
Lou - Yes, my understanding is that it does depend on antibodies being produced. You asked what the LFA antigen is doing in this type of test. I don't understand why you are asking since from what I can tell, it is measuring antibodies to an antigen but then I am just learning about all of this so maybe you can explain why you ask please?
According to the info from Immunosciences from the link that Bea sent:, "Simultaneous detection of antibodies against OspA and its cross-reactive epitope in human tissue (LFA) may identify individuals with treatment-resistant arthritis. It may also provide a predictive model for the development of autoimmune inflammatory disease induced by Borrelia." I also found this information here: Entrez PubMed
Scott - 1) If you have symptoms, these tests seem to support chronic infection in my opinion.
The problem is that according to the text that came with the test, one cannot be considered positive for lyme if they have a response to the "other spirochete". They also state that one must have a reaction to at least one of the 3 specific Borellia antigens. That seems wrong to me since there are 100 strains and they are only testing for 3 but then maybe there is something else involved that I don't understand.
3) Immunosciences test is NOT a good test for coinfections. I would get coinfection tests done elsewhere. They only test for one stain of each, thus potentially missing some of the strains that are also common, especially the Ehrlichias.
Thank you for mentioning this. I didn't know. I just recently had IgeneX babesia FISH (RNA), Babesia WA-1 serology IFA, Microti antibody G/M, HME panel (monocytic), Henselae antibody G/M, MDL bartonella species panel by PCR, Babesia microti by real time PCR, babesia microti real time by ELISA and they were all negative.
I personally find the test misleading in that they call it a test for "Coinfections" when in reality it is really not. They are looking for cross-reactivity with Lyme, but not really attempting to do a true coinfection test.
Yes, I see your point. Hopefully my doctor will be able to tell something from all these negatives and one positive on a test that isn't even really meant to test co-infections.
Treepatrol - thanks for the information. In the last article you gave. "This combinatorial variation could potentially produce millions of antigenic variants in the mammalian host."
I'm sure that I don't completely understand the implications of this but it seems like that alone would make testing via antigens a hit and miss situation however I don't enough about this to know just how different the antigens can get or if they would be different enough to NOT produce an immune system response against a certain antigen even though one was originally infected with that antigen. Hope that makes sense. LOL
When I find out anything, I'll add it to this thread. My phone appt with my LLMD isn't for 2 weeks but I think he is pretty savvy with these tests. Hopefully I'll know appropriate questions to ask him by then. Terry
Posts: 6286 | From Oregon | Registered: Jan 2006
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SForsgren
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That is frustrating that they will not provide more specific information. That is one of the challenges I have found with Immunosciences tests is that they are often so complex (maybe advanced actually) that the doctors don't know how to read them either. I bet you would not find a doctor that knew the answer to the question you posed. Try also sending an email to the email on their web site and just start off
Dr. Vojdani...
and see what happens. I have met Aristo a few times and he has always been very gracious in answering questions.
I hope that your coinfection test results are really the case. Personally, I never believe that someone has Bb and no co-infections. It may take time for them to appear, but the odds are very much against it.
A 2.4 for Babesia likely is indication that you have had exposure to Babesia in the past even though they don't test for all strains. You at least know B microti has been there from what it appears.
Did you do an IgeneX WB as well? I find that doing more than one test and cross referencing is helpful.
-------------------- Be well, Scott Posts: 4617 | From San Jose, CA | Registered: Jul 2005
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TerryK
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Thanks for your suggestions Scott. I'll try writing and see if that get's me anywhere. I'll let you know if it does, but again, I won't hold my breath. You obviously have had better luck than I have because I have never been able to talk to anyone that knows anything at any of the labs.
I'm thinking that my doctor may be better equipped than many to deal with the technical aspects of some of these lab tests because I've heard that he has family in the tick testing business. That may not be accurate but that's what I've heard. I'll add to this thread after I talk to him in early June.
I have had an IgeneX IgG and IgM back at the end of 2003. IgM had 3 equiv bands (31, 39, 41), IgG was IgeneX positive but one band short of meeting CDC criteria. Positive bands were (18, 39, 41, 58), equiv (28, 45, 66, 93). IgG ELISA was equiv., IgM ELISA was neg. I was told by my doctor (not an LLMD) that I didn't have lyme.
I was pretty sick at the time but after a few months decided to start researching lyme and eventually started samento which I took for 7 months. Now, 6 months after going off samento I got an IgM from IgeneX which currently shows postive bands for 30, 58 and 83-93. Equiv bands are 39 and 41. I did not get another IgeneX IgG but did get an MDL IgM and IgG recently. MDL states IgM, no bands present, IgG alt is Equiv with the following bands present (60, 41, 39 and 23).
In light of the Lyme serology from immunosciences, it seems possible that I am infected with some unknown spirochete that my immune system identifies as Bb or of course it could be some strain of Bb. In any case, I and many of my family members are infected with something because we are all very sick with lyme and babesia like symptoms. I plan to seek aggressive treatment and see what happens.
I've read many of your responses and you always provide thoughtful input and I thank you for the time you spent on this one as well. Terry
Posts: 6286 | From Oregon | Registered: Jan 2006
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TerryK
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I had the phone appointment with my LLMD this week and I want to report back with the limited information that I have about this test.
I faxed my questions before the appointment, two of which were the questions that I initially posed in this thread.
1. Are there any other known infections that cause similar antigens and can therefore cross react? If so, what are they?
2. What, are the implications that I tested above the range for "unknown spirochete"? a. Does this imply that my immune system may be causing me to erroneously test positive on some of the Western Blot bands that are specific to Bb?
In my LLMD's opinion, the fact that I tested positive for "unknown spirochete" does not have any implications. Spirochetal infections are not unusual, an example he gave was dental spirochetes.
My LLMD said that the fact that I showed protiens to Borellia was significant and that this test was different than the Western Blot. He didn't say this but I assume that means that this test is another verification besides the Western Blot of the presence of borellia.
Sorry to say that we didn't discuss the test any further because we only had 15 min to cover a LOT of ground as there were many other tests to discuss.
Combining the results of this test with my other tests, helped to determine the liklihood of a lyme diagnosis. We did not discuss the fact that babesia showed up on this test. I had quite a few other babesia tests that showed up negative but I will bring this up with him in my next appointment since I know that babesia is another one that does not have accurate tests yet.
I have an office appointment at the end of July and I plan to take these questions and get further info then. I will report back when I have further information.
I never got a call back from immunosciences. I did not write to them. They said that the doctor that Scott mentioned does not talk to patients so I don't feel it is worth the trouble.
On the liklihood of lyme - My doctor said that the positive 83-93 kDa result on the IgeneX IgM is significant since it is 92-94% specific for Borellia.
In his opinion I have lyme but the diagnosis is not a slam dunk.
I am having another IgM next week after being on treatment for a month to see if further bands show up.
BTW - At the end of 2003 I had an IgeneX positive IgG with 4 positive bands.
I think this is interesting because it illustrates that lyme is a clinical diagnosis and even though we may have bands showing that are Bb specific, that is not absolute positive proof that we have lyme. OTOH, having no bands showing is not proof that we don't have lyme. sigh
I know there are a lot of opinions about the interpretation of tests and the liklihood of lyme diagnosis and who knows who is right but this LLMD is well respected and I feel that he is being appropriately cautious.
When I first saw him, he told me that based on my symptoms and my IgeneX positive IgG Western Blot (not CDC positive but IgeneX positive), there was a 70-80% and above chance that I have lyme. The latest tests make it more likely that I have lyme and we are doing more tests to further narrow the liklihood.
I am remaining open minded because *if* borellia and co's are not causing my symptoms, I want to know so that I find out what is and fix it. Personally, I think Bb or some other similar spirochete and/or co-infection are likely at the root of my and my family's symptoms and so I am being treated. My response to treatment will provide more information.
My doc said that even though anti-virals are said to be ineffective for the herpes viruses, he has seen some people improve on them and that is an option down the road if I don't get better with only lyme treatment. I'm also looking into elevated lead levels and food allergies from the results of my testing. Terry
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