Borreliosis serology testing complete & honest study

Topics with information and discussion about published studies related to Lyme disease and other tick-borne diseases.
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LHCTom
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Borreliosis serology testing complete & honest study

Post by LHCTom » Sat 18 Jul 2015 11:37

Have you ever wanted to a study that doesn't leave out 80% of possibilities or make assumptions with no apparanet agenda?

A neutral European, Swedish study not biased by US politics or dogmas

Good history summary and ideas for algorithms more sensible than 2nd grade 2 tiered.

Genospecies/strain impcat on serology
Bb, Bg, Ba may vary at IR6 region by as many as 6 Amino Acids impacting C6 ELISA

Testing
IFSA
ELISA
Western Blot
Culture
PCR
lymphocyte transformation test

Time from bite
Strain/species
Bb
Ba
Bg

versus

Virotech (VT) Borrelia burgdorferi ELISA IgG/IgM test kit (REV 031127-05)
Virotech Borrelia burgdorferi ELISA
Diasorin Liaison (Li) Borrelia IgM (310890), IgG (310880)
IR6 peptide antibody analyses
Dako Cytomation IDEIA Lyme neuroborreliosis test kit K6028 (IgM and IgG anti-flagellum antibodies)
Virotech Borrelia Ecoline IgG/IgM Line Immunoblot (WB EL ) (REV 040202-01)
The following antigens were used for IgM antibodies: OspC, VlsE and borrelial membrane protein A (BmpA or p39)
For IgG antibodies the following antigens were included: VlsE, BmpA, p83, BBA36, BBO323, Crasp3 and pG.
Plus EBV capsid antigen gp125 for cross reaction control
R&D Systems Quantikine Human CXCL13 Immunoassay (DCX130)

versus

acrodermatitis chronicum atrophicans (ACA)
Lyme arthritis (LA)
Borrelial lymphocytoma (BL)
Lyme neuroborreliosis (LNB)
Lyme carditis (LC)
Erythema migrans (EM) at initiation
Erythema migrans (EM) + 2-3 months
Erythema migrans (EM) + 6 months
Blood donors ( presumably healthy)
Age <= 12 >12 years of age

with

WB banding as appropriate
Band profile of WB
Cross reacting with:
Epstein Barr virus (EBV)
Rheumatoid Factor (RF)
Borrelial lymphocytoma (BL)
Treponema pallidum
syphilis serology RPR and TPPA test

and

Innate immune response pathogenesis
Th1/Th2 shifts
T & B cell response
Serum
Spinal Fluid (CSF)
Immunoglobulin as we age
Two tiered versus single tier
Possible history of Blood Donor healthy controls past Borrelia exposure increaes with age

http://www.diva-portal.org/smash/get/di ... TEXT01.pdf

Found on Google search "borrelia recombinant monoclonal antibodies C6 elisa ppt"

I consider this the way real scientists design and execute studies when they feel free to publish openly without peer review back pressure to prove dogmas.

Its a long read but well worth it!
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

duncan
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Joined: Wed 5 Sep 2012 18:48

Re: Borreliosis serology testing complete & honest study

Post by duncan » Sat 18 Jul 2015 12:26

LHCTom, wow, I want to review this but the link isn't working for me.

When I key in your search combination, a scrolling list of PDF files appear, and my brain cannot figure out which one is the right one.

It's me, isn't it? I am the only one that could not figure this out.

Sigh.

dlf
Posts: 294
Joined: Sun 7 Apr 2013 15:36

Re: Borreliosis serology testing complete & honest study

Post by dlf » Sat 18 Jul 2015 23:54

LHCTom, many thanks for posting this. It will take me awhile to go through it thoroughly, but on a first look it is very interesting!
I would really like to track down the antigens used in some of those tests. While several are listed, others are not. According to the study they used a version of the Immunetics C6 that was based on the IR6 of Borrelia garinii. In North America the Immunetics C6 peptide ELISA uses Borrelia burgdorferi strain B31 recombinant protein, so I am guessing they must produce and sell a different version of the C6 for the European market. I think we need to be careful to not confuse the two different versions.

Duncan, you must have used LHCTom's suggested search. He did include the actual link in red. It works just fine for me. I will post it as a link, but in the future when you see a url and it isn't linked, hold your left mouse button down at the front of the url and move it to the end before releasing the button. That area should then turn blue on your screen showing you have selected it. Put the cursor somewhere in that blue area and right click. It will ask you if you want to go to.......left click on the line that asks that and away you should go directly to it. Here is the link:

http://www.diva-portal.org/smash/get/di ... TEXT01.pdf

Some of the other papers that are the foundation for this one are also interesting.

LIST OF PAPERS
I. I Tjernberg, G Krüger, I Eliasson: C6 peptide ELISA test in the
serodiagnosis of Lyme borreliosis in Sweden, Eur J Clin Micorbiol Infect
Dis. 2007 Jan. 26(1):37-42.
I think this was the first one and it is abstract only. After that they started using diva-portal for full text distribution.

http://www.ncbi.nlm.nih.gov/pubmed/17180348

Paper 2,. I Tjernberg, T Schön, J Ernerudh, AC Wistedt, P Forsberg, I Eliasson:
C6-peptide serology as diagnostic tool in neuroborreliosis, APMIS. 2008
May. 116(5):393-9.

Sorry I didn't look this one up.

3. III. I Tjernberg, H Sillanpää, I Seppälä, I Eliasson, P Forsberg, P Lahdenne:
Antibody responses to borrelia IR(6) peptide variants and the C6 peptide
in Swedish patients with erythema migrans, Int J Med Microbiol. 2009
Aug. 299(6):439-46.
http://www.diva-portal.org/smash/get/di ... TEXT01.pdf

4. I Tjernberg, AJ Henningsson, I Eliasson, P Forsberg, J Ernerudh:
Diagnostic performance of cerebrospinal fluid chemokine CXCL13 and
antibodies to the C6-peptide in Lyme neuroborreliosis. J Infect. 2011 Feb.
62(2):149-58.
http://liu.diva-portal.org/smash/get/di ... TEXT01.pdf

I also found some other interesting material by I Tjernberg:

Ivar Tjernberg, Martin Carlsson, Jan Ernerudh, Ingvar Eliasson and Pia Forsberg
Mapping of hormones and cortisol responses in patients after Lyme neuroborreliosis

BMC INFECTIOUS DISEASES, 2010, 10(20), .

http://liu.diva-portal.org/smash/get/di ... TEXT01.pdf

And one more by a co-author on Paper 4 on Immunological aspects of Lyme: (which probably should be used to start a new thread, but I haven't done that).

Doctoral thesis, comprehensive summary (Other academic)
Clinical, Epidemiological and Immunological Aspects of Lyme Borreliosis
with Special Focus on the Role of the Complement System
Anna J Henningsson
http://www.diva-portal.org/smash/record ... swid=-6550

http://www.diva-portal.org/smash/get/di ... TEXT01.pdf

duncan
Posts: 1370
Joined: Wed 5 Sep 2012 18:48

Re: Borreliosis serology testing complete & honest study

Post by duncan » Sun 19 Jul 2015 13:53

Thanks, dlf. I think I finally got it.

Assuming I did, I agree, there is a wealth of diagnostic possibilities. I'd like to dig further into the IR6 and similar stuff.

I did find some of it to be a genetic outgrowth of sorts from the IDSA school, though. Lots of questionable assumptions, and references to peeps like Steere and Wormser. For instance, the author just assumes there is no b garinii or afzelii in North America, and as many of us have pointed out: Pretty much no one is looking for either species in the Western Hemisphere. Also, he seems to willingly subscribe to special characterizations, e.g. Sensu stricto is characterized by Lyme arthritis, and very rarely, NB...

Perhaps I am misunderstanding. It IS a long read.

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LHCTom
Posts: 341
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Re: Borreliosis serology testing complete & honest study

Post by LHCTom » Sun 19 Jul 2015 20:02

Duncan,

You have to keep in mind this is a thesis paper done by a Swedish Graduate student who just finished reading all the textbooks and research papers and so he can't help but have a medical literature bias since one exists throughout ID medicine. That being said, he was young and very thorough trying to impress his pfofessors and the others involved in his studies. The important thing is he classifies everyone so well and takes into account possible past Borrelia exposure and so many variables we have been screaming about for so long and in a honest questioning tone. He points out the Bb. Ba and Bg all have different epitopes so their are really at least 3 C6 tests, one for each invariable section 6 ( and is this extrapolate to all species and some strains?)

He explains the tests he uses for the ELISA and discusses the antigens used and sonicate versus recombinant ( made by cloning), and his list of antigens by the suppliers don't even come close to the CDC magic 10 (VlsE, BmpA, p83, BBA36, BBO323, Crasp3 and pG.). Why?

He discusses cross reactivity and how using the EBV gp125 antigen can be used on the blot as an antigen to specifically exclude EBV.

He is afraid to mention infection can take as short as 24hours.

He isn't afraid to mention that different strain and species that have nucleotide errors can cause changes in eptitopes that reduce or eliminate epitope to antigen binding and by implication B31 could cause this.

He discusses the various main initial symptom of each sttrain Ba, Bb, Bg and isn't afraid to explore antibody performance at EM EM+2-3 months and 5 months. Would be nice if it included 1 and 5 years but studies must finish before he graduates.

He reviews cross reacting and shows WB profiles and discusses everthing from Th1/Th2 shifts and their characteristic cytokines and Il-x

He is afraid to mention the probaability as we age of having a past Borrelia exposure in controls.

He mentions immunoglobulins impact somewhat.

Look at his tables at the eend and show me one US researcher who reviews so many compounding hypothesis and shows so much data per patient.

One can tell he is young with some dogmas already imbedded but his naivity make him more creative and open than I've ever seen..

Read it carefully and look at the tables in the back - its different...

Its a perfect example of how youth without too many assumptions and the use of honest creativity emerges when you haven't been beaten into the group position yet. :evil: :? :shock: :lol: :oops: :mrgreen: :| :bonk:

I''m not saying its perfect but its refreshing to see so many Lyme issues touched upon when US researchers like Wormser wouldn't dare say any of the the "it could have been caused by" or "this should be considered" or "the C6 has at least 3 variants due to amino acid changes due to species" or "dare to mention antigens used by test supplier", or "oldre blood donor healthy controls may have had Borrelia exposure as a functionof age", or dare to mention " one tier versus 2 tired", or give antibody profiles over "0, 2-3 and 6 months" or "mention time to infection from bite" etc...

Think about all the maybe statements he makes that are reasonable and would never be made here.
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

duncan
Posts: 1370
Joined: Wed 5 Sep 2012 18:48

Re: Borreliosis serology testing complete & honest study

Post by duncan » Sun 19 Jul 2015 20:37

Understood. Thanks for setting me straight. Sometimes I can't see the forest...

I will study those tables and the other areas you and dlf noted more closely. I've already bookmarked it.

X-member
Posts: 3954
Joined: Mon 30 Jul 2007 18:18

Re: Borreliosis serology testing complete & honest study

Post by X-member » Fri 24 Jul 2015 15:28

Maybe this belong in this thread?

Laboratoriediagnostik av borreliainfektion
(Laboratory diagnosis of Lyme borreliosis)

SMI (Infectious Diseases Institute), Sweden

https://www.folkhalsomyndigheten.se/pag ... 101-28.pdf

A (not perfect) google translated quote:
The German Society of Lyme disease (Borreliose Deutsche Gesellschaft, 2010), which is a independent association, believes that all serological diagnosis of BI must be made with WB since only then can assess antibody specificity profile.
Despite the new specific primary screening tests using recombinant antigens as VlsE or C6 peptide, they think also that with only ELISA faces up 15 percent false negative samples.
BI = borrelia infection

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