Monday, March 2, 2009

The amazing Lyme WB 100 Kd band???

Clongen labs reports the whole spectrum of Lyme WB bands. The Bands as seen on a Western Blot strip are shown heaviest to lightness. Other labs that I am familiar with report the heaviest band as 83-93. Lyme WB strips from Clongen show "all the bands"; there are more than 14. The Clongen strips frequently show reactivity at 100 Kd band which exceeds the control response. I asked Dr. K what this 100 Kd band was. He did not know. He just presents the data. Intrigued, I did a little research. I have found nothing in human literature. But I did find something.

Of particular interest is a paper called: Lyme-Disease- "a consensus statement"
This paper is talking about canine, not human Lyme disease.

The piece was written in 2005. It comes from a German lab- LABOKLIN.
The bands discussed in the paper are for the most part quite familiar. They include:
the 24Kd band (23-25), the 31Kd band, the 39Kd band and the 41Kd band.
The article notes that the 41 band is associated with flagellin protein and that the 31 band is associated with OSPA. Sound familiar?

They paper claims that the best Lyme WB band is the 100 Kd band. Although it is seen only in late disease, the authors state "...there is not known cross reaction and (it) is therefore pathognomonic." The means that reactivity at the 100 Kd WB bands confirms Lyme exposure with 100% accuracy. If the same holds true in humans this may be incredibly significant!

Comments please!

9 comments:

Leslie said...

Interesting research, doc. Now, if research would be done for humans....

Thanks for your hard work and enlightenment!

girl mark said...

I'd read somewhere that veterinary lyme testing is better than that for humans. I'd assumed it was due to differences in our biology. I'd love to hear DogDoc's comments on that one. I'm writing this comment from my phone so I can't easily Google what the veterinary tests actually are.

is it actually just an issue of reporting?

joysie said...

As one of your patients with a positive 100, I am anxious to hear comments and further discussion on this. How many people show a positive 100?

gimpy said...

I'm also curious how many of your patients who show 'only a 41 band' or one of the other supposedly controversial bands and are otherwise considered 'seronegative' but have have the 100 Kd band- how long has Clongen been reporting the 100 Kd?

This is the kind of data that would be useful for someone to mine out of LLMD's records (assuming that's legal to do)- something like a Turn The Corner grant like what is paying for research into patterns found in Dr Burrascano's patient records?

gimpy said...

sorry about the typo or lack of clarity- the question was "do you know yet how many of your patients who tend to turn up as 'only a 41 band' actually have both a 41 and a 100Kd?"

Lyme report: Montgomery County, MD said...

There is no clear definition of seronegative or seropositive.

For many LLMDS a single 23,31,34,39 or 93 band is seropositive. How the 100 band may add to this is unknown.
Other LLMDS consider a 41 band to be suggestive. Some consider IgeneX IND bands to be significant. Some doctors consider a C6 peptide index of 0.3 or greater to suggest seropositivity. Basically the focus on seronegativity and seropositivity may be misplaced. Largely, Lyme remains a clinical diagnosis.

dogdoc said...

I beleive the article you are referring to is discussing the results of the recombinant antigen western blot for dogs in used commonly in Europe and one common product is marketed through that laboratory. If this is the case, the 100kd band they are referring to is for the p100 Borrelia antigen. This is considered by most in humans and animals to be a specific band and to occur late in neuroborreliosis. From my understanding, p100 can have three variations and can migrate to 83kd, 93kd, or 100kd position depending on the different borrelia species. In Europe, it usually shows at 83 or 100 and they combine those two positions as one band in there interpretation of human western blots. It can be a bit more confusing in the newer recombinant line blots that use the recombinant p100 antigen which don't really report the weight of migration like the standard blots we are used to. Here are just a couple of references- it could be that we have a european species here in our patients that we are not looking for. Or that the p100 protein in our local strain has the 100kd varient instead of the 83kd or 93kd ones.
www.pubcentral.nih.gov/articlerender.fcgi?aritd=1233984$tool = pmcentrez Improvement of Lyme Borreliosis Serodiagnosis by a Newly Developed Recombinant IgG and IgM Line Immunoblot Assay and Addition of VslE and DbpA Homologues Goettner, et al J Clin Microbiol 2005, August 43(8) : 3602- 3609

www.pubmedcentral.nih.gov/articlrender.fcgi?artid + 86736 & tool = pmcentrez A European Multicenter Study of Immunoblotting in Serodiagnosis of Lyme Borreliosis Robertson, et al J Clin Microbiol 2000 June; 38 (6) 2097-2102

PatJ. said...

The shampoo of my dogs reads: "when your dog has fleas and ticks your family and pets may be exposed to: - Lyme disease, tapeworm infestation, flea bite anemia and flea allergy dermatitis"? See that it does not say dogs contaminated by Deer’s fleas... so, does this means these diseases have been ignored in humans? might it also mean that labs for pets are aware of the infection and can detect it better, easier and are not limited to just look for the Lyme in the "fleas of the Deer"? Do they have the accurate tests?
(Other discussion, some "regular fleas" from dogs or cats or some "regular ticks" from dogs or cats might also transmit the disease?)

Dear Dr. in Montgomery, this "discovery" you have made, of the 100 band, might save many from lots of suffering. Hopefully Labs will be able to detect Lyme in humans more precisely and guidelines from these organizations will have to open to treat the disease even after when they think they already treated it and it is a post- syndrome!

My question is, how to make a regular Doctor, or a "Lyme specialist", to get to do this test? the ones I've met just do the Elisa, after much begging and do not even know how to interpret it!

Love my dogs!

Pseudonymity said...

As a supplement to the comments by dogdoc, from http://www.ncbi.nlm.nih.gov/pubmed/8282192

The p100 gene coding for the p100 protein of Borrelia burgdorferi strain PKo has been cloned, sequenced and expressed in Escherichia coli. An open reading frame including upstream and downstream sequences with potential translation and transcription signals could be identified. The reading frame consists of 1989 nucleotides corresponding to a protein of 663 amino acids and a calculated molecular mass of 75.8 kDa. The protein has a leader peptide and is processed without modification at the N-terminus. A high percentage of amino acid sequence identity could be found to the high-molecular mass protein p83/p93 of B. burgdorferi strain B31.

It looks like the distinction between 83, 93 and 100 kDa locations on the WB is probably strain dependent. Note the referenced publication is from 1993 - recombinant p100 was available even before Dearborn.

I think that for experienced chronic Lyme patients, the fact that not all bands are equal is a pretty well known fact. The strain variation of p100 also does not say much for the standard commercial blot kits in the US, which I understand are typically based on B31 only.