-------------------- --Lymetutu-- Opinions, not medical advice! Posts: 96222 | From Texas | Registered: Feb 2001
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bpeck
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posted
You'd be positive by European standards. You'd be positive by US reference standards (IGENEX)
But you'd be negative by USA CDC criteria.
Barb
PS. Strongly positive on Band 39 Kda is considered positive for Lyme by anyone who knows anything about Lyme. That band is not cross reactive with anything else, and highly specific for Lyme.
Posts: 1875 | From VT | Registered: Oct 2002
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posted
If these results are positive by European standards then someone forgot to tell Scotland that they are part of Europe! My husband has these results and his test has been considered equivocal. Do you know where I can find guidance papers on European standards?
Posts: 72 | From Scotland | Registered: Apr 2005
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bpeck
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posted
Well I guess there's as big of a disconnect with the European researchers and the Practicing Clinicians in Europe as there is in the USA.
The following references are what I thought was the European Criteria which was adopted.
So what IS the testing criteria?
It can't be USA's Dearborn criteria- because your borrelia strains are different.
Barb
Clin Microbiol. 1997 Jun;35(6):1433-44. Interpretation criteria for standardized Western blots for three European species of Borrelia burgdorferi sensu lato.
Hauser U, Lehnert G, Lobentanzer R, Wilske B.
Max von Pettenkofer-Institut fur Hygiene und Medizinische Mikrobiologie der Ludwig-Maximilians-Universitat Munchen, Munich, Germany.
Western blots (WBs; immunoblots) are a widely used tool for the serodiagnosis of Lyme borreliosis, but so far, no defined criteria for performance, analysis, and interpretation have been established in Europe. For the current study WBs were produced with strains PKa2 (Borrelia burgdorferi sensu stricto), PKo (Borrelia afzelii), and PBi (Borrelia garinii). To improve resolution we used gels of 17 cm in length. In a first step, 13 immunodominant proteins were identified with monoclonal antibodies. Then, the apparent molecular masses of all visually distinguishable bands were determined densitometrically. Approximately 40 bands of between 14 and 100 kDa were differentiated for each strain. From a study with 330 serum samples (from 189 patients with Lyme borreliosis and 141 controls), all observed bands were documented. To establish criteria for a positive WB result, the discriminating ability of a series of band combinations (interpretation rules) were evaluated separately for each strain (for immunoglobulin G [IgG] WB, > 40 combinations; for IgM WB, > 15 combinations). 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. The overall sensitivity was the highest for PKo WB, followed by PBi and PKa2 WB, in decreasing order. Standardization of WB assays is necessary for comparison of results from different laboratories.
Validity of interpretation criteria for standardized Western blots (immunoblots) for serodiagnosis of Lyme borreliosis based on sera collected throughout Europe.
Hauser U, Lehnert G, Wilske B.
Max von Pettenkofer-Institut fur Hygiene und Medizinische Mikrobiologie der Ludwig-Maximilians-Universitat Munchen, D-80336 Munich, Germany.
Western blotting (WB; immunoblotting) is a widely used tool for the serodiagnosis of Lyme borreliosis (LB), but so far, no generally accepted criteria for performance and interpretation have been established in Europe. The current study was preceeded by a detailed analysis of WB with whole-cell lysates of three species of Borrelia burgdorferi sensu lato (U. Hauser, G. Lehnert, R. Lobentanzer, and B. Wilske, J. Clin. Microbiol. 35:1433-1444, 1997). In that study, interpretation criteria for a positive WB result were developed with the data for 330 serum samples (from patients with LB in different stages [n = 189] and from a control group [n = 141]) originating mostly from southern Germany. In the present work, the interpretation criteria for strains PKo (Borrelia afzelii) and PBi (Borrelia garinii) developed in the previous study were reevaluated with 224 serum samples (from patients with LB in different stages [n = 97] and from a control group [n = 127]) originating from throughout Europe that were provided by the European Union Concerted Action on Lyme Borreliosis (EUCALB). De novo criteria were developed on the basis of the reactivities of the EUCALB sera and were evaluated with the data for the samples from southern Germany. Comparison of all results led to the following recommendations: For WB for immunoglobulin G (IgG), at least two bands among p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for PKo and at least one band among p83/100, p39, p30, OspC, p21, and p17b for PBi; for WB for IgM, at least one band among p39, OspC, and p17 or a strong p41 band for PKo and at least one band among p39 and OspC or a strong p41 band for PBi. WB with PKo was the most sensitive, and this strain is recommended for use in WB for the serodiagnosis of LB throughout Europe.
Publication Types: Multicenter Study
PMID: 10364592 [PubMed - indexed for MEDLINE
Posts: 1875 | From VT | Registered: Oct 2002
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bpeck
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Member # 3235
posted
Up for Brodie
Posts: 1875 | From VT | Registered: Oct 2002
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posted
Just for interest, although I'm in Scotland and not in the USA, I emailed CDC yesterday and asked them why, when p39 is a specific band for borellia, they still demand other bands to call the test positive. Their response says
"Some people with exposure to bacteria related to the bacterium that causes Lyme disease may have factors in their blood that cross react on western blot causing a false positive test. This is why we can not rely on one or two strongly positive bands".
So does this mean that specific bands are not specific? Or is it just a get out clause?
bpeck
Frequent Contributor (1K+ posts)
Member # 3235
posted
Gee- as long as they've started a dialogue on this topic- email them back and ask to see the references which show cross reactivity with Band 39 kDa. they could also include any references to molecular mimicry, autoimmunity - or other pathogens.. or even tissue type for that matter.
I have never seen reference that ANYTHING cross reacts with Band 39 kDa.
posted
Ok Barb - I've asked them for specific references on 39kDa. This is the third time I have got them to answer questions - I seem to get a response within 24 hours every time. I write like I am a grateful idiot who knows nothing and who thinks they are wonderful - and they fall for it every time. It will be interesting to see if they still do so when I start to get a little more technical! If it still works, I'll get more technical still! They will probably get fed up with me soon, especially when they must know from my email address that I am not in the USA.
posted
Well, I asked CDC for a specific reference for cross-reactions to 39 kDa - and boy did I get no answer. I quote:
"Finding information about specific proteins is complication by nomenclature. The 39 kDa protein of Borrelia burgdorferi belongs to the Bmp protein family. A great resource for researching medical publications is called PubMed. PubMed is a service of the National Library of Medicine and includes over 14 million citations for biomedical articles back to the 1950's. You can access PubMed on the Internet at the following address:"
Or in other words, find it yourself - if you can. Sorry Barb - end of enquiry I think.
posted
When I posted my husband's WB result, I thought this was the result of the test done about 7 months after he was bitten. BUT I've just discovered this is the result of a WB they did on the blood sample drawn only 3 weeks after the bite. So, would the CDC not count this as positive since it was so early on? It seems to fit their 2 out of 3 by being strongly positive at 39 kDA and 41 kDA. If so, I can point out to his ID Consultant that he would have been positive in the USA even if not positive in Scotland.
Posts: 72 | From Scotland | Registered: Apr 2005
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posted
Up for responses to new question. See above.
Posts: 72 | From Scotland | Registered: Apr 2005
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bpeck
Frequent Contributor (1K+ posts)
Member # 3235
posted
If 39 and 41 are positive on an IgM western blot, then yes it would be positive even by CDC standards.
But I assumed we've been talking about an IgG Blot as your conversation has cenetered around the requirement for more bands to be present to be considered positive.
Barb
PS- if these are the results of an IgG western blot, and he had no IgM bands present - but had band 39 and 41 present on an IgG blot, then he's probably been bitten in the past.
IgM antibodies are formed first, and they live about 3 months - there's an over lap when the IgG antibodies are formed so after about 3 months or so it's IgG that remain to do the killing and IgM are usually no longer present.
But we know in Lyme- IgM can spike up any time Lyme comes out into the blood - so IgM can go up and down over time.
Here's M. Kaplans site on Lyme- alot of your questions can be answered here:
posted
I don't know whether it was an IgM or IgG blot that was done - but I can't see the lab doing an IgG when told it was only 3 weeks since the bite. I'll follow that up anyway. But I think I'll still point out to the Consultant that these bands constitute a positive IgM in the USA.
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