Please see the message below in reference to the presentation by Isabella W. Martin, MD,  Assistant Professor of Pathology and Laboratory Medicine, Geisel School of Medicine, Dartmouth

---------- Original Message ----------
From: CARL TUTTLE <runagain@comcast.net>
To: William Marsh <wmarshmd@gmail.com>, CHAIR
Cc: All Members of the Study Commission
Date: 04/01/2021 8:24 AM
Subject: Re: Topic for discussion at our next meeting (Dr. Martin's Presentation #2)

To the Chair and members of the Lyme Study Commission,
 
I would like to call attention to the following statement from Dr. Martin’s presentation:
 
@ 24:03 “Direct detection of the organism itself is not particularly useful”
 
https://www.youtube.com/watch?v=kfRWPPp_KW0
 
 
Carl Tuttle’s comment:
 
Culture is the gold standard for definitive diagnosis of bacterial and fungal infections worldwide but when it threatens the existing paradigm suddenly it becomes “not particularly useful.”
 
Persistent Borrelia Infection in Patients with Ongoing Symptoms of Lyme Disease
http://www.mdpi.com/2227-9032/6/2/33
 
All patients in this 2018 Middelveen et al pilot study were culture positive for infection (genital secretions, skin and blood) even after multiple years on antibiotics so there was no relief from current antimicrobials. Some of these patients had taken as many as eleven different types of antibiotics.
 
 
Culture medium used:
 
 Barbour-Stoner-Kelly H (BSK) complete medium with 6% rabbit serum (Sigma-Aldrich, #B8291, St. Louis, MO, USA)  [1]
 
-To avoid contamination, all cultures were performed under strict aseptic conditions in a laboratory that was free of  Borrelia reference strains, and cultures of control and patient samples were processed in an identical manner.
 
- Borrelia DNA in extracted samples was detected using a published TaqMan assay targeting a 139-bp fragment of the gene encoding the Borrelia 16S rRNA, as described previously [59,63,64]. Amplifications were conducted on a CFX96 Real-Time System (Bio-Rad, Hercules, CA, USA) with cycling of 50 °C for 2 min, 95 °C for 10 min, followed by 40 cycles of 95 °C for 15 s and 60 °C for 60 s, and fluorescent signals were recorded using CFX96 Real-Time software with the Cq threshold set automatically.
 
- PCR amplification was followed by Sanger sequencing. PCR products were extracted using the QIAquick Gel Extraction kit (Qiagen, Hilden, Germany) in accordance with the manufacturer’s instructions. Eluates were sequenced in both directions, then were compared by BLAST analysis using the GenBank database (National Center for Biotechnology Information).
 
 
Carl Tuttle’s comment:
 
This is why patients remain sick indefinitely while our public health officials turn a blind eye to the evidence that we are dealing with an antibiotic resistant/tolerant superbug and instead of creating a Manhattan Project to find a cure they regurgitate the disinformation year after year decade after decade.
 
 
Carl Tuttle
Hudson, NH
 
Reference
 
 
1 Standardization of medium for culturing Lyme disease spirochetes.
 
R J Pollack, S R Telford, 3rd, and A Spielman
 
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC262913/
 
The standardized medium is designated BSK-H