The following study is important to patients because:

Molecular Detection of Borrelia in Human Tissue

Hasibul Haque – Dalhousie University, Halifax, Nova Scotia

Summary of Slide presentation:

  • 52 known species of borrelia
  • 21 members of the Lyme group
  • 29 members of relapsing fever group

At 2:28 shows Bb can be spirochetes, round bodies, blebs, and biofilm.  Persister form is highly resistant to treatment

Molecular methods 

  • enables direct detection at onset of clinical signs
  • identifies type of borrelia
  • detects borrelia and coinfections in particular sample
Hypothesis:

Molecular detection of biopsy, necropsy and autopsy tissue for Borrelia and other associated bacteria will allow us to understand the relation of borrelia infection to the associated disease(s).

Objective:  Evaluate the value of direct molecular detection of borrelia in human tissues.

Targeted tissues:  synovial membrane, major vasculature, nervous tissue, kidney etc.

Testing used:  Nested PCR (amplifies DNA), Molecular beacon, and immunostaining.    

Info for BB3 (patient used for study)

Study:  

  • 70 year old male with history of Lyme confirmed by 2 tier Canadian Serology
  • Received standard treatment (21 days) of oral antibiotics
  • Cause of death was severe coronary artery disease with associated extensive atherosclerosis
Type of samples from BB3

Ethanol submerged tissue (pericardium, endocardium aortic valve, aorta, liver, pancreas cerebral cortex)

  • tested by nPCR and found positive for flagellinB.  Confirmed by sequencing.  Primers OspA, Flagellin B, 235 Burgdorferi

Molecular beacon from BB3 showed 3 slides at 9:53

  • Slide A:  positive control for Bb (mouse brain 400X magnification)
  • Slide B:   BB3 FISH of Bb in endocardium sample from patient 400X magnification which is positive for Bb
  • Slide C:  negative control for Bb in mouse brain, 400X magnification

parafin embedded tissue

Immunostaining for BB3  showed round body (persister form), spirocheteal form

  • Slide A positive control for Bb in mouse liver (spirochetal, 
  • Slide B Bb found in endocardium sample from patient
  • Slide C negative control for Bb in mouse liver

Conclusions:

  • Bb found in ALL targeted tissue samples
  • Molecular beacon shows round bodies & spirocheteal forms
  • Immunostating also shows Bb in different forms
  • standard treatment did not eliminate Bb

Summary:

  • Shows value of molecular detection of Bb in human tissues
  • Findings are consistent with tick exposure and serology
  • Molecular detection offers the possibility of determining if Bb in present
  • Offers opportunity to address other research questions such as borrelia distribution & correlation with tissue damage.

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**Comment**

We need more work like this done and less on blood serology which continues to turn up seronegative decade after decade.

The form of testing used to determine Lyme infection has been a source of heated debate from the beginning:  https://madisonarealymesupportgroup.com/2018/04/03/cdc-deliberately-avoids-direct-detection-testing-methods-for-ld/  Excerpt:

It would appear that there has been a deliberate avoidance of direct detection methods and it is believed that these efforts are to insure that the current thirty year dogma remain intact.

We have a dire need to develop rapid detection methods for a serious growing health threat which has the ability to disable its victim as described in the attached letter addressed to the previous Director of the CDC. (Please see attachment in link)

I would like to point out that employees of the U.S. Centers for Disease Control hold patents on metabolomics (Lyme tests).

CDC Employee Patent: https://www.google.com/patents/EP2805168A1?cl=en

For nearly four decades now the only FDA approved test for Lyme disease is the indirect two-tiered antibody test. Direct detection methods to identify the causative agent responsible for the disease have been avoided, criticized and shelved.

https://madisonarealymesupportgroup.com/2018/12/16/laboratory-testing-for-lyme-disease/  Direct detection laboratory testing (DNA/PCR Sequencing) is used for many infections (Ebola (1), Zika (2), Bartonella (3) etc.) but not Lyme disease.

More on testing:  https://madisonarealymesupportgroup.com/2018/09/12/lyme-testing-problems-solutions/

https://madisonarealymesupportgroup.com/2018/10/13/direct-test-for-ld-carl-tuttle-chews-up-cdc-spits-them-out/

https://madisonarealymesupportgroup.com/2017/12/13/suppression-of-microscopy-for-lyme-diagnostics-professor-laane/  Excerpt:

After publishing the 2013 article ‘A simple method for the detection of live Borrelia spirochetes in human blood using classical microscopy techniques’, professor Laane was invited to give a lecture at the 2014 Norvect conference in Oslo. An English patient saved the pdf, so you can still read it, via the link provided.

I was present at that conference and still remember how nervous he was. The reason was that several medical professors complained to his university. He was threatened with losing his job, if he would speak at the conference.

In fact, he did not literally speak – as you can see in the movie below – but used performing arts to show the slides of the spirochetes. Professor Laane was fired anyway and his laboratory was closed down.