Improved Borrelia burgdorferi Blood culture Method

Medical topics with questions, information and discussion related to Lyme disease and other tick-borne diseases.
Pandora
Posts: 252
Joined: Tue 20 Mar 2012 14:58

Re: Improved Borrelia burgdorferi Blood culture Method

Post by Pandora » Thu 15 Aug 2013 22:04

http://jcm.asm.org/content/early/2013/0 ... M.01674-13
Assessment of New Culture Method to Detect Borrelia species in Serum of Lyme Disease Patients
Barbara J.B. Johnson#,
Mark A. Pilgard and
Theresa M. Russell
- Author Affiliations
Division of Vector-Borne Diseases, Centers for Disease Control and Prevention Fort Collins, CO, USA

ABSTRACT

A novel method of culturing spirochetes from the serum of U.S. Lyme disease patients was recently reported by Sapi and colleagues to have 94% sensitivity and 100% specificity for Borrelia species as assessed by microscopy and DNA sequence analysis of the pyrG gene (Sapi E, Pabbati N, Datar A, Davies EM, Rattelle A, Kuo BA. 2013. Int J Med Sci 10:362-376). The majority of the spirochetes described were related by pyrG sequences to species of Borrelia previously undetected in North American patients without a reported travel history to Europe or Asia. To better understand these unexpected findings, we determined pyrG sequences of the laboratory reference strains used by the investigators for method development and testing of culture medium.

Eighty percent (41/51) of the reported patient-derived pyrG sequences are identical to one of the laboratory strains and an additional 12% (6/51) differ by only a single nucleotide across a 603bp region of the pyrG gene. Thus, false positivity due to laboratory contamination of patient samples cannot be ruled out and further validation of the proposed novel culture method is required.
-------------------------------
Ummm tell me why CDC does not unnerstand Antigenic variation?
http://www.biomedcentral.com/1471-2180/9/137
Genome sequencing of diverse bacterial species has revealed widespread distribution of conserved gene products

with as-yet unknown functions.

Among these are a family of small proteins with approximate molecular masses of 12 kDa, which have been variously classed as

"domain of unknown function" (DUF) 149, Pfam 2575 and COG-0718 [1].

Such genes have been identified in a wide variety of bacterial phyla,

a list that includes many significant pathogens of humans, domestic animals and plants (Fig. 1).

thumbnailFigure 1. Alignment of the predicted amino acid sequences of YbaB/EbfC orthologs of
++++++++++++++++
H. influenzae (Hi),
++++++++++++++++
E. coli (Ec),

Vibrio cholerae (Vc),

Pseudomonas putida (Pp),
Rickettsia rickettsiae (Rr),
Neisseria gonorrhoeae (Ng),
Bdellovibrio bacteriovorus (Bba),
Clostridium perfringens (Cp),
Bacillus subtilis (Bs),
Enterococcus faecalis (Ef),
Streptococcus pneumoniae (Sp),
Mycobacterium tuberculosis (Mt),
Bacteroides capillosus (Bc), and

B. burgdorferi (Bbu).

Identical amino acids are boxed and shaded. Amino acid residues of YbaBEc and YbaBHi that comprise αlpha-helices 1 and 3 of their determined protein structures are identified.
DNA-binding by Haemophilus influenzae and Escherichia coli
------------------------
Bpur, the Lyme disease spirochete's PUR-domain protein: identification as a transcriptional modulator and characterization of nucleic acid interactions.
http://www.ncbi.nlm.nih.gov/pubmed/23846702
J Biol Chem. 2013 Jul 11. [Epub ahead of print]

Jutras B, Chenail AM, Carroll DW, Miller MC, Zhu H, Bowman A, Stevenson B.
Source

University of Kentucky, United States;
Abstract

The PUR-domain is a nucleic acid-binding motif

found in critical regulatory proteins of higher eukaryotes,

and in certain species of bacteria.

During investigations into mechanisms by which the Lyme disease spirochete controls synthesis of its Erp surface proteins,

it was discovered that the borrelial PUR-domain protein,

Bpur, binds with high affinity

to double-stranded DNA

adjacent to the erp transcriptional promoter.

Bpur was found to enhance the effects of the erp repressor protein,
BpaB.

Bpur also bound single stranded DNA and RNA,

with relative affinities RNA > double stranded DNA > single stranded DNA.

Rational site-directed mutagenesis of Bpur identified amino acid residues

and domains critical for interactions with nucleic acids,

and revealed that the PUR-domain has a distinct mechanism of interaction

with each type of nucleic acid ligand.

These data shed light on both gene regulation in the Lyme spirochete

and functional mechanisms of the widely-distributed PUR-domain.
----------------------
CDC is under the Authority that if they don't find IT-IT does NOT EXIST!

Pandora
Posts: 252
Joined: Tue 20 Mar 2012 14:58

Re: Improved Borrelia burgdorferi Blood culture Method

Post by Pandora » Thu 15 Aug 2013 22:09

And I'll go ya one better...just for fun...
http://www.the-scientist.com/?articles. ... -Proposed/

http://en.wikipedia.org/wiki/File:Elect ... A_H7N9.png
Better grab that one if you want it before its gone...

http://www.biomedcentral.com/1471-2180/9/137
Genome sequencing of diverse bacterial species has revealed widespread distribution of conserved gene products

with as-yet unknown functions.

Among these are a family of small proteins with approximate molecular masses of 12 kDa, which have been variously classed as

"domain of unknown function" (DUF) 149, Pfam 2575 and COG-0718 [1].

Such genes have been identified in a wide variety of bacterial phyla,

a list that includes many significant pathogens of humans, domestic animals and plants (Fig. 1).

thumbnailFigure 1. Alignment of the predicted amino acid sequences of YbaB/EbfC orthologs of
++++++++++++++++
H. influenzae (Hi),
++++++++++++++++

http://www.youtube.com/watch?v=H29IBRlFn-M
http://www.youtube.com/watch?v=pqKaM_J7KDI
http://www.youtube.com/watch?v=1ojq_2-HlNg

And ya wonder how in the world did they get so stupid?
Well they did infect all they could for decades....
http://www.youtube.com/watch?feature=pl ... Ono_2m_8LY

edbo
Posts: 90
Joined: Sat 2 Feb 2013 21:48

Re: Improved Borrelia burgdorferi Blood culture Method

Post by edbo » Fri 16 Aug 2013 1:48

Pandora wrote:Thus, false positivity due to laboratory contamination of patient samples cannot be ruled out and further validation of the proposed novel culture method is required.
Wow, that's a first step! To me this looks like a great success when the CDC is looking at Sapi et al's paper and trying to validate it (or rather invalidate it) instead of all the other lyme "experts" outright ignoring the findings.

Can anyone translate for me what Johnson is saying? Is she saying that the strains that Sapi considered unknown are in fact identical to the laboratory strains?

And what are they doing to try to (in)validate the new culture method? Are they running it themselves? Then it would be easy for them to check laboratory contamination...

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inmacdonald
Posts: 977
Joined: Fri 13 Jan 2012 22:32

Re: Improved Borrelia burgdorferi Blood culture Method

Post by inmacdonald » Fri 16 Aug 2013 14:00

CDC Critique of Sapi et al Blood Culture for Borrelia in Human Blood:Many Mistakes by the CDC
critique methods and False Conclusions:
_________________________________________________________________________
CDC Analysis of the DNA sequencing Data for the Advanced Labs Blood culture isolates:
by Dr. Barbara Johnson et al.
________________________________________________________________________
An Official CDC critique and an Official CDC accusation of alleged Flawed Advanced Laboratory techniques
resulting in Only 3 genovars instead of 60 + Genovars of Borrelia as reported by Sapi
__________________________________________________________________________
The CDC Analysis is flawed for the reasons posted below and the CDC manuscript must be
withdrawm
_________________________________________________________________________

The Reader of the CDC manuscript will infer that The CDC had in their possession
aliquots of each of the isolates from  Advanced Labs
If this is NOT the case.


The CDC may be required to issue a Retraction of theirmanuscript
to clarify this point for the reader and to address the
BLASTn data which follows for a single isolate .
Advanced Laboratories and the CDC agreed had 21 nucleotide mismatches with B31 were present
in the exemplary Isolate. [Isolate JX867376}


CDC analysis of isolate JX86 7376  detected 21 base mismatches with respect to B31
    and the Sapi Paper also reported 21 base mismatches with respect to B31 [97% identity with B31]
I did a BLASTn search on isolate  JX867376 against all borrelia.

A 100% match was obtained with JX867376 itself as deposited in GenbBank.This is expected
and this 100% match indicates that the 603 bases were correctly entered into the NCBI BLASTn
Search Engine.
-------------------
  B31 borrelia complete DNA sequence produced a result with 21 mismatches against Sapi deposit JX867376 for a congruence percentage of 97%
  It is interesting that the BLASTn search also revealed 97% matches for the additional Borrelia on Deposit in GenGank:
          N40, CA382 ZS7 and multiple numbered haplotypes of Borrelia spp.
   A 96% match was noted for multiple genotypes of B. garinii and B. Bavariensis.

So if the truth be told, based on a BLASTn search of all Borrelia in GenBank, it
   is very very misleading to state that isolate JX867376 was UNIQUE for B31 
    as is posted at position 49 in the CDC Table.....AND .....

and to omit any mention of the following borrelia genotypes
   which also yielded 97% matches with JX867376 according to BLASTn search.
ST12
N40
ST8
CA382
ST4
ST51
ST56
St1
ZS7
Each and every one of the above genotypes should have been posted at postion 49 in the CDC table.
To post only B31 is very misleading to the reader who does not use the BLASTn tool
for sequence analysis and borrelia genotyping.
______________________________________________
Correction to the CDC Table of Results for (97%) [21 base mismatches against reference strain borrelia burgdorferi B31] against Advanced Labs Isolate JX867376.

" This Advanced labs isolate may indicate ANY of the following Borrelia species:
B31, N40,ZS7,ST! ST4, ST8ST12,,ST51,ST56, CA382,
_______________________________________
_______________________________________

Mismatches of Bases by BLASTn analysis should now be initiated for Each and Every Advanced Lab nucleotide sequences
   and the original manuscript should be withdrawn and re-written
    to accommodate BLASTn search data for each of the Advanced Lab  PyrG nucleotide sequences.


I look forward to CDCreconciliation of BLASTn data with the observed Advanced Lab dep[osited nucleotide Sequences
   with the hope that the issue of Contamination of Patient Recovered Borrelia from blood cultures
     may be viewed in the full context of BLASTn search Data  to embrace all possible borrelia species with
       closely matching nucleotide sequences.

The maximum number of polymorphic sites in CDC analysis of the Sapi Data was 21.
CDC speciated only 3 strains of borrelia in Their analysis :
B31 burgdorferi [N=22], 
B023 afelii [N=2}, and 
Fuji P1 garinii [ n=27]
These assignments conflict with the Sapi paper as follows:
In the Sapi manuscript Table 3 :
Sapi et al reported
5 isolates of Garinii - [ no identical garinii genotypes noted by Sapi }
1 isolate of afzelii Pko [consistent with European type afzelii]
3 kurtenbachii  [ all genotypically distinct]
1 americanum and
6 burgdorferi [B31=1] , [ZS7=1} , [ N40=1], { ST1 =1], { ST11 =1}, { St8=1}.

For the remaining: "JX86***** series isolates I will consult the GenBank accessions
as I have for isolate JX867376....
and perform my own Clustal analysis. of nucleotide Base alignments

CDC group registered  the following genovars [ your results in parentheses]
among the JX86**** series:
7424 51 polymorphic sites   [Called Afzelii B023 by CDC group]
7398 49 polymorphic sites    [called Afzelii B023 by CDC group]
7394 43 polymorphic sites     { Not classified by CDC group}
7417 42 polymorphic sites    [Called Garinii Fuji P1 by CDC group}
7380 42 polymorphic sites    {called Garinii Fuji P1 by CDC group}
7378 42 polymorphic sites     { Called Garinii Fuji P1 by CDC group}
7379 42 polymorphic sites      Called Garinii Fuji P1 by CDC group}

Twenty one isolates with 41 polymorphic sites EACH [ isolates not collated here]
7393 39 polymorphic sites  [Called garinii Fuji P1 by CDC group}
7376 21 polymorphic sites    [[See above - called B31 burgdorferi by CDC group}}
7419 2 polymorphic    sites    [ Called burgdorferi B31 by CDC group}
 
CDC also indicate that CDCS equence analysis yielded
27 Identical isolates of Garinii
2 i dentical isolates of Afzelii. 
22 isolates of B31 { with only 2 cases #48 and 49 called B31 burgdorferi with "mismatched" bases : #48 n=1 miusmatch and #49 with 21 mismatches]
So instead of 60+ unique Borrelia genovars, the CDC reduced the Sapi/Advanced Labs results to just 3 genovars
-----------------------------------------------------
I insist the the genotyping of the Advanced Labs GenBank deposits
speaks for heterogeneity of genovars, and reinforces the concept that So called 
European type borrelia may very well be now recognized as vectored by USA Ixodid ticks.
Mexico, Central America and Brazil are geographies in which GARINII has been proven
to be vectored by ticks.  To make matters more complex, the Amblyomma family of ticks (Amblyomma cajenesse)
are competent vectors for Garinii infections in the Western Hemisphere.
We need not look to Sea bird migrations to explain USA cases of Ixodid scapulairs
and Amblyomma spp. vectored human infections.

The uniqueness of the Sapi report of Advanced Labs Human Blood culture Borrelia strains
is therefore CORRECT.
No Laboratory Contamination of patient material with any living borrelia ever occurred
European Strains of borrelia were recovered from the blood of USA patients in the
Advanced Labs study of Borrelia blood cultures

Respectfully submitted,
Alan B. MacDonald MD FCAP FASCP
August 16,2013

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LHCTom
Posts: 341
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Re: Improved Borrelia burgdorferi Blood culture Method

Post by LHCTom » Sat 17 Aug 2013 1:56

Accordingly, we determined the relevant portion of the pyrG DNA sequences of the other laboratory strains and deposited them in GenBank [KF170280 (strain BO23); KF170281 (strain 297); and KF170282 (strain Fuji P1)
The CDC pyrG DNA sequences are not on Genbank. Other Burgdorferi strains have the exact same or close pyrG sequence. This means the set of isolates near or matching B31 really means nothing. I will provide specifics from Blast.

Its not possible to confirm that the isolates match the strain BO23 or strain Fuji P1 pyrG DNA sequences until the CDC actually deposits them in GenBank. Why would these not be deposited prior to publication. I was unable to find them in GenBank either from previous submissions or the claimed CDC submission. Fuji p2 is available but is not meaningful.

Even if there is a close match between the BO23 strain and the isolates, it will require a thorough review of all strains with similar 603 sequences to know if it means real contamination. Based on a brief Blast review, there are multiple other pyrG sequences that are identical and many at 99%. I will provide a summary of the other possible genotypes from Genbank.

So in order to determine if the CDC paper conclusion is meaningful - Its necessary to:

1) Get access to the CDC pyrG DNA sequences for strain BO23 and strain Fuji P1
2) Blast Search all 603 bp pyrG DNA sequences to see what the other possibilities are
3) If they are only the CDC sequences, then that is worthy of concern
4) If the 603 bp pyrG DNA sequences are common like B31, then the conclusion is meaningless

Lets wait for the CDC to publish the sequences before making any conclusions. The CDC should have deposited the sequences and provided a thorough review of the pyrG variations across all known/published 603 kb sequences. Without this, its simply not possible to conclude anything. I hope they are not trying to pull the wool over our eyes? They wouldn't do that would they? no?


If you accept the CDC arguments, then all 51 isolates were contamination. If this were true, then how did the 48 controls avoid contamination? The CDC would like you to believe they escaped because they were not subject to DNA analysis. But the CDC claims half were contaminated with B31?? Why were they not caught by the monoclonal antibody? The CDC avoids this and claims the B. garinii or B. afzelii were missed by the Bb monoclonal antibody. But the pyrG gene is not adequate to exclude Burgdorferi as the species.

The CDC weaves a nice sounding story until you look closely. Lets see the CDC pyrG sequences and we can take it from there....

Nice try but very sloppy!
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

Pandora
Posts: 252
Joined: Tue 20 Mar 2012 14:58

Re: Improved Borrelia burgdorferi Blood culture Method

Post by Pandora » Sun 18 Aug 2013 23:27

THANK YOU BRAVE KNIGHTS!!! WE WERE HOPEING YOU WOULD SAY THAT!!!

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LHCTom
Posts: 341
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Re: Improved Borrelia burgdorferi Blood culture Method

Post by LHCTom » Mon 19 Aug 2013 2:13

I’ve been reviewing the culture using the NCBI BLAST program and it doesn’t look good. BLAST is an online program that allows searching of all the NCBI Genbank DNA sequences for similarity to either one of the already submitted sequences by Accession Number or a user sequence. The ALS paper has 20 identical Borrelia garinii sequences and 5 with one character and two with 2 and 3 differences (CGAT). Since garinii should be unusual or rare in the US, having 25 + 5 + 1 + 1 = 27 = more than 1/2 is a concern. So I looked closer. I checked whether any non-garinii or burgdorferi pyrG genes were close to the garinii and there were none close =99%. They all differed by quite a few characters. That means these samples were almost certainly garinii. Then I checked the 5 with one difference and discovered they all=5 had one character difference from the 25 but they were random = none the same.

That suggests they were most likely errors in the sequencing and not 5 different variants. Therefore all 25 were very likely the same strain. I then checked if there were any other garinii pyrG sequences that matched. The 2 that matched were from Japan and Russia - see below example. The garinii JujiP1 used by ALS was also from Japan. I found it in a 1995 paper. The CDC has yet to submit their sequences to Genbank. There were others with a one character difference which were also from Japan. So the genotype found in the culture 27 times was almost certainly garinii probably from Japan or possibly Russia – not the US nor even Europe. Not good!

This is still a mystery since the controls had ZERO contamination as seen by the monoclonal or polyclonal antibodies??? Its a shame ALS didn't PCR the pyrG gene in the controls. What are the odds, these controls would have gotten through the same culture process without a single contaminant? The burgdorferi and even the afzelii results seem plausible but the 27 garinii are very troubling. If the garinii pyrG genes all varied significantly, that would suggest garinii has been missed in the US. But they don't.

Sorry, but I feel compelled to tell it like I see it.

Here is the BLAST link

You can enter Accession numbers in the box - run BLAST and it will list all similar sequences. Then you can look at each of them compared to determine "how many different" and a link to its description.

http://blast.ncbi.nlm.nih.gov/Blast.cgi ... =blasthome

example:

JX867375.1 - one of the garinii isolates

There are 603 characters in most pyrG genes

100% = 603/603

At 100% =exact there are 2 exact matches:

Borrelia garinii strain Ekb704-11 CTP synthase (pyrG) gene, partial cds GenBank: JX971327.1
Borrelia garinii pyrG gene for CTP synthase, partial cds, isolate: HP1 GenBank: AB555778.1

For 99% there is a long list that are all garinii

98% starts with a few garinii followed by:

Borrelia bavariensis strain PZwi CTP synthase (pyrG) gene, partial cds GenBank: KC833620.1 = 592/603 = 11 characters

94% is where the first appears after bavariensis 6% is about 40 characters difference

Borrelia bissettii pyrG gene for CTP synthase, partial cds GenBank: AB526143.1
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

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inmacdonald
Posts: 977
Joined: Fri 13 Jan 2012 22:32

Re: Improved Borrelia burgdorferi Blood culture Method

Post by inmacdonald » Mon 19 Aug 2013 21:34

A CDC published paper with Lies and flawed Analytics; the Borrelia Blood Culture Critiques; 2013 August 14
RETRACTION of the published paper is now indicated

The CDC authors , have:
1. Lied to the readers of J.Clin Micro--because they did not place into the GenBank
three of the alleged :
KF170280 [ allegedly derived from burgdorferi sl strain B023]
KF170281 {allegedly derived from burgdorferi ss. strain 297}
KF170282 [allegedly derived from burgdorferi sl strain Fiji{1]
2. Lied to the readers when they wrote that living borrelia from strain B023 and from strain FijiP1
were the source of Contamination of human blood culture specimens.


3.Lied to the readers of the J. Clin Microbiology when they stated that Garinii type borrelia infections
have never been reported in the Western Hemisphere
, In spite of previously published manuscripts
in the CDC publication "Emerging Infectious Diseasees" with a report of 2 Borrlelia lymphocytoma cases
in patients from Mexico [ EID ,2007,oct,13(10):1556-8], One case of borrelia lymphocytoma from Midwestern USA
Wis Med J. 1990 Dec;89(12):683-6.
Borrelia lymphocytoma: a possible North American case.
Finkel MF, Johnson RC.
SourceWestern Wisconsin Lyme Disease Center, Midelfort Clinic, Eau Claire, WI 54701.,
and in spite of manuscript reports from Brazil implicating Borrelia garinii
as an Etiologic agent of Brazilain Lyme borreliosis.{Talhain,S. et al, borrelia burgdorferi SLin Brazil: Occurrences confirmed by IHC FFM", Acta Tropica: 115(3) 200-4, and Pirana, S. et al," sero-reactivity to Bb, BAzelii and B. Garinii antigens in patients afected by peripheral facial palsy in Brazil" ,Otology and Neurology, 2002,(23)S33.

4. Failed to use a simple freely publically available software [Clustal w for DNA sequences alignments} to
critically examine the "same-ness' or the "different-ness" of the Advanced labs pyrG DNA sequences
which were posted on the National GenBank site for public examination.
5. Failed to understand that in addition to Garinii type FujP1 --- the Advanced Labs group also utilized
5 additional Garinii type strain pyrG sequences in their Quality control procedures -
with DNA sequences for Ab555778.1,Jf331269,JF331231,Jf331248,Jf331265
included in their scientific study.
that adds up to 6 Garinii Controls on the Advanced Labs side and one [Unpublished ??isit even in existence??]
Garinii control on the CDC side.
Who did more complete "Due Diligence" in scientific controls?
6. The final two sentences of the CDC paper state:
" Independent verification is particularly critical when claims are at odds with a larg body of other scientific work and when they may trigger unnecessary antibiotic treatment ot patients"'
______________________________________________________________________________________________
MacDonald's comment and challengeto the CDC: [their sentence in quotation marks aboe the line}
Where is the documentation that there was is or shall be "un-necessary antibiotic treatment in any of the
patients who received a positive result from Advanced labs. Did patients with a negative Advanced Labs report
for detection of borrelia inbloodcultures - did any of these person opt for no further antibiotic treatment
for borrelia or for any of themany co-infections which are agreed are co-travellers with borreliosis?_______________________________________________________________________________________________
"We Caution clinicians and patients to wait for independent verification by Scientifically sound methods before using this culture service for diagnostic services"
_______________________________________________________________________________________________
MacDonald's comment and challenge to the CDC: [ their second sentence in quotation marks above the line]
Which of the Authors of the CDC paper has a license to practice medicine?
Is the CDC now in the practice of medicine. It is malpractice for a physician to prescribe treatment
for patients whom he/she has never seen nor personally examined.
Were the CDC scientific methods "sound" in the rendering of their paper in J. Clin .Microbiology
doi:10.1128/JCM.01674-13 ?
Were any of the intricate laboratory methods used by Advanced Laboratories in the least bit
"scientifically UNSOUND????"
Guilt for lack of Sound science falls squarely on the shoulders of the authors of the CDC
in their article doi 10:1128/JCM.0164-13. Retraction of this article is the only acceptable
next step by Johnson, Figard, and Russell of the CDC Fort Collins Co.USA facility.

REspectfully,
Alan B.MacDonald, MD
August 19,2013

User avatar
inmacdonald
Posts: 977
Joined: Fri 13 Jan 2012 22:32

Re: Improved Borrelia burgdorferi Blood culture Method

Post by inmacdonald » Mon 19 Aug 2013 21:34

A CDC published paper with Lies and flawed Analytics; the Borrelia Blood Culture Critiques; 2013 August 14
RETRACTION of the published paper is now indicated

The CDC authors , have:
1. Lied to the readers of J.Clin Micro--because they did not place into the GenBank
three of the alleged :
KF170280 [ allegedly derived from burgdorferi sl strain B023]
KF170281 {allegedly derived from burgdorferi ss. strain 297}
KF170282 [allegedly derived from burgdorferi sl strain Fiji{1]
2. Lied to the readers when they wrote that living borrelia from strain B023 and from strain FijiP1
were the source of Contamination of human blood culture specimens.


3.Lied to the readers of the J. Clin Microbiology when they stated that Garinii type borrelia infections
have never been reported in the Western Hemisphere
, In spite of previously published manuscripts
in the CDC publication "Emerging Infectious Diseasees" with a report of 2 Borrlelia lymphocytoma cases
in patients from Mexico [ EID ,2007,oct,13(10):1556-8], One case of borrelia lymphocytoma from Midwestern USA
Wis Med J. 1990 Dec;89(12):683-6.
Borrelia lymphocytoma: a possible North American case.
Finkel MF, Johnson RC.
SourceWestern Wisconsin Lyme Disease Center, Midelfort Clinic, Eau Claire, WI 54701.,
and in spite of manuscript reports from Brazil implicating Borrelia garinii
as an Etiologic agent of Brazilain Lyme borreliosis.{Talhain,S. et al, borrelia burgdorferi SLin Brazil: Occurrences confirmed by IHC FFM", Acta Tropica: 115(3) 200-4, and Pirana, S. et al," sero-reactivity to Bb, BAzelii and B. Garinii antigens in patients afected by peripheral facial palsy in Brazil" ,Otology and Neurology, 2002,(23)S33.

4. Failed to use a simple freely publically available software [Clustal w for DNA sequences alignments} to
critically examine the "same-ness' or the "different-ness" of the Advanced labs pyrG DNA sequences
which were posted on the National GenBank site for public examination.
5. Failed to understand that in addition to Garinii type FujP1 --- the Advanced Labs group also utilized
5 additional Garinii type strain pyrG sequences in their Quality control procedures -
with DNA sequences for Ab555778.1,Jf331269,JF331231,Jf331248,Jf331265
included in their scientific study.
that adds up to 6 Garinii Controls on the Advanced Labs side and one [Unpublished ??isit even in existence??]
Garinii control on the CDC side.
Who did more complete "Due Diligence" in scientific controls?
6. The final two sentences of the CDC paper state:
" Independent verification is particularly critical when claims are at odds with a larg body of other scientific work and when they may trigger unnecessary antibiotic treatment ot patients"'
______________________________________________________________________________________________
MacDonald's comment and challengeto the CDC: [their sentence in quotation marks aboe the line}
Where is the documentation that there was is or shall be "un-necessary antibiotic treatment in any of the
patients who received a positive result from Advanced labs. Did patients with a negative Advanced Labs report
for detection of borrelia inbloodcultures - did any of these person opt for no further antibiotic treatment
for borrelia or for any of themany co-infections which are agreed are co-travellers with borreliosis?_______________________________________________________________________________________________
"We Caution clinicians and patients to wait for independent verification by Scientifically sound methods before using this culture service for diagnostic services"
_______________________________________________________________________________________________
MacDonald's comment and challenge to the CDC: [ their second sentence in quotation marks above the line]
Which of the Authors of the CDC paper has a license to practice medicine?
Is the CDC now in the practice of medicine. It is malpractice for a physician to prescribe treatment
for patients whom he/she has never seen nor personally examined.
Were the CDC scientific methods "sound" in the rendering of their paper in J. Clin .Microbiology
doi:10.1128/JCM.01674-13 ?
Were any of the intricate laboratory methods used by Advanced Laboratories in the least bit
"scientifically UNSOUND????"
Guilt for lack of Sound science falls squarely on the shoulders of the authors of the CDC
in their article doi 10:1128/JCM.0164-13. Retraction of this article is the only acceptable
next step by Johnson, Figard, and Russell of the CDC Fort Collins Co.USA facility.

REspectfully,
Alan B.MacDonald, MD
August 19,2013

hv808ct
Posts: 256
Joined: Wed 30 Jul 2008 4:11

Re: Improved Borrelia burgdorferi Blood culture Method

Post by hv808ct » Tue 20 Aug 2013 15:17

A CDC published paper with Lies and flawed Analytics; the Borrelia Blood Culture Critiques; 2013 August 14
RETRACTION of the published paper is now indicated

The CDC authors , have:
1. Lied to the readers of J.Clin Micro…
2. Lied to the readers…
3. Lied to the readers of the J. Clin Microbiology…
Right. Journal authors routinely lie and papers not meeting the approval of Internet-based amateurs should be retracted upon demand. The only thing “indicated” here is that: 1) it’s never too late to take a typing class and to learn how not to capitalize or use a bold font in random, rambling fashion, and 2) hysterical typing and accusations may lead to misreadings and misstatements as in the following example.

“Lied to the readers of the J. Clin Microbiology when they stated that Garinii type borrelia infections have never been reported in the Western Hemisphere, In spite of previously published manuscripts in the CDC publication "Emerging Infectious Diseasees" with a report of 2 Borrlelia lymphocytoma cases in patients from Mexico [EID ,2007,oct,13(10):1556-8], One case of borrelia lymphocytoma from Midwestern USA Wis Med J. 1990 Dec;89(12):683-6. Borrelia lymphocytoma: a possible North American case.”

Better re-read the EID article (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2851501/) and note the word “possible” in that 23-year-old Wisc. Med. Article.


Of more interest, however, is the original paper by Sapi et al., the end of which acknowledges the following: “This work was supported by a grant from RAM Capital LLC to the Research Division of Advanced Laboratory Services Inc (ALSI).”

Advanced Laboratory Services is located at 501 Elmwood Avenue, Sharon Hill, PA 19079 (which may be the same thing as Hometech Therapies, Inc. of 501 Elmwood Avenue, Sharon Hill, PA 19079). (website: http://www.hometech-rx.com). ALSI also sounds a lot like Advanced Research Corporation based in Florida.

Company Overview of Advanced Research Corporation.
Advanced Research Corporation, a contract research organization, provides clinical research and consultation services for pharmaceutical, biotechnology, and medical device industries. Advanced Research Corporation was incorporated in 2000 and is based in St. Petersburg, Florida at 100 2nd Avenue South, Suite 401, St. Petersburg, FL 33701. Founded in 2000. Phone: 727-897-9115; Fax: 727-897-9095.

Key Executives for Advanced Research Corporation
Dr. Avery Huff
Chief Operating Officer
Dr. Joseph J. Burrascano
Senior Vice President of Medical Affairs and Medical Director
Mr. Stephen Duprez
Director of Clinical Operations
Ms. Donna Fordham
Project Director
Ms. Phyllis Clark
Project Director

But who is RAM Capital?

Ram Capital Group LLC in Sharon Hill, PA is a private company categorized under Investors (Unclassified). Our records show it was established in 2005 and incorporated in Pennsylvania.
Ram Capital Group, LLC
501 Elmwood Avenue
Sharon Hill, PA 19079-1014
There is no Key Executives data available.

Just that same Elmwood address. Anyone ever been there?

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