The Primers Shell Game, a version for the Facebook "Occupy the DOJ" group, a chapter of CRYME DISEASE and a lesson from the new “Khan Academy” of Lyme Cryme and HLA-Negative Diseases (includes all abused groups; CFIDS, ME, FM, Autism, medical rights, parents rights, psych rights, etc); 141031, KMD.
Okay, let me repeat, this is a chapter of “CRYME DISEASE.” Which means there is more than one chapter of CRIME DISEASE, and the primary chapter is Chapter One. Chapter One is about Scientifically Valid Testing for Lyme (which is not arbitrary but follows FDA rules), which includes this same information, below, that you need to understand about the genetic relatedness of these kinds of spirochetes. Another chapter is 7 (BRAIN_PERMANENT.htm), where the crooks almost always use the OspA gene instead of the flagellin gene or a species-specific spacer gene to find "NO LYME," so you have to compare the following data to that, that is, if you want more examples than the following...
Flagellin is the species distinguisher.
All Borreliae are relapsing fever organisms, and the nature of the relapse is antigenic variation. Therefore you cannot use any DNA from plasmids – which is where the variable surface antigens are ordered manufactured and remanufactured – to assess for spirochetes. No one sane does this. No researchers outside the United States EVER uses plasmid DNA to assess for spirochetes. They only use species-specific spacer genes like 5, 16 and 23 S RNA or flagellin. That’s it.
Let’s start at the beginning because the moron “LLMDs” out there are killing me, really. I feel like I am in Beatrice’s World. (“That’s not how this works.”)
Now, let's side-step for a minute and talk about the FDA and what their rules are for the "Validation of an Analytical Method." As you can see there is Accuracy (should detect 100% of the instances when the analyte in question is present), Specificity (only detects one thing), Linearity, Ruggedness, Precision (refers to instrumentation), Limit of Detection (this would be something like, "How low in concentration of the analyte in question can your method detect?").
This is from the new announcement July 31, 2014 regarding the FDA now about to ENFORCE their validation rules:
Do you see the FDA wrote their words here? I underlined them.
"Sensitivity" MEANS "Limit of Detection." The closest thing to that in the real VALIDITY requirements is Limit of Detection.
FDA Rules on the VALIDATION of an Analytical Method
Specificity (only detects one thing)
Accuracy (Should detect 100% of the instances where the analyte is present, and the concentration should be close to 100% of that known to be spiked in, and never should detect "none" as is the case with Lyme Western Blotting and the Lyme ELISA, especially)
Limit of Detection (means "What is the lowest concentration of the analyte in question does your method detect?")
Precision (system has integrity in performance)
Ruggedness (anyone can run the test with their own equipment and get the same results)
Linearity (concentration range of analyte for which the test is valid in and out of matrix or "inert ingredients")
But first, your test should detect all the cases in question, - or be 100% ACCURATE - and that means, in the case of Lyme, the only analyte that can be tested for is flagellin or anti-flagellar antibodies. Anti-flagellar antibodies can be found in probably 95% of Lyme cases. So, Yale went ahead and made that Specific (US patent 5,618,533) in 1991, here, next:
Says Yale: "Molecular characterization of the humoral response to the 41-kilodalton flagellar antigen of Borrelia burgdorferi, the Lyme Disease spirochete."
"The earliest humoral response in patients infected with Borrelia burgdorferi, the agent of Lyme disease, is directed against the spirochete's 41-kDa flagellar antigen. In order to map the epitopes recognized on this antigen, 11 overlapping fragments spanning the flagellin gene were cloned by polymerase chain reaction and inserted into an Escherichia coli expression vector which directed their expression as fusion proteins containing glutathione S-transferase at the N terminus and a flagellin fragment at the C terminus. Affinity-purified fusion proteins were assayed for reactivity on Western blots (immunoblots) with sera from patients with late-stage Lyme disease. The same immunodominant domain was bound by sera from 17 of 18 patients. This domain (comprising amino acids 197 to 241) does not share significant homology with other bacterial flagellins and therefore may be useful in serological testing for Lyme disease."
Okay, so Yale says there, ▲ above, that their method (US patent 5, 618, 533) detects, early, late, neurological, and every other possible kind of Lyme outcome and that it detects 94.4% of the cases, which means it is the closest possible method we could possibly have to detect Lyme ("should be 100% of the cases," says the FDA, verbatim), and this method was made SPECIFIC, which means it does not detect any other flagellins.
When the FDA says "sensitivity," they really mean "LIMIT OF DETECTION" and refers to the METHOD and not the "CASES." Accuracy addresses cases. Yale took care of all that in 1991 and went ahead and patented it (but did not use this method to qualify LYMErix, their other patent, which is the essence of this False Claims Act case).
Okay, so in summary >> The only way to detect a spirochetal disease is to use recombinant specific flagellins from most of the specific borreliae - just like Yale did, above, only with the other borrelia - that we know to be at least in the United States. THAT is what is "VALID," and the FDA and NIH agree with me.
Okay, so: DO NOT EVER listen to IDSA, the Lymedisease.org (LDA) or ILADS.org. IDSA are criminals and the LDA/ILADS are retarded and have no real scientists in their camps.
ILADS and the LDA do not know what they are talking about and consistently come up with stupid, retarded announcements, giving everyone with Lyme-brainscramble and everyone who is an MD but has no science background... even more brainscramble. It just is not fair.
If you don't know what you are talking about, kindly do not talk.
Single Spirochete Infection and resultant MULTIPLE VARIANTS:
1951: Relapse Phenomena in Rats with a Single Spirochete
Oscar Felsenfeld and CDC officer Alan Barbour talking about/referencing this Single Spirochete Phenomena:
Oral Spirochetes infecting Alzheimer’s brains and traveling along inside nerves (this is not the only report that says this, you’ll find it in syphilis report too) (from BRAIN_PERMANENT.htm); An independent study on spirochetes in the brain from dentists and they say:
Molecular and immunological evidence of oral Treponema in the human brain and their association with Alzheimer's disease.
Riviere GR, Riviere KH, Smith KS.
Department of Pediatric Dentistry, School of Dentistry, Oregon Health and Sciences University, Portland, OR 97201-3097, USA.
“The purpose of this investigation was to use molecular and immunological techniques to determine whether oral Treponema infected the human brain. Pieces of frontal lobe cortex from 34 subjects were analyzed with species-specific PCR and monoclonal antibodies. PCR detected Treponema in 14/16 Alzheimer's disease (AD) and 4/18 non-AD donors (P < 0.001), and AD specimens had more Treponema species than controls (P < 0.001). PCR also detected Treponema in trigeminal ganglia from three AD and two control donors. Cortex from 15/16 AD subjects and 6/18 controls contained Treponema pectinovorum and/or Treponema socranskii species-specific antigens (P < 0.01). T. pectinovorum and/or T. socranskii antigens were also found in trigeminal ganglia and pons from four embalmed cadavers, and 2/4 cadavers also had Treponema in the hippocampus. These findings suggest that oral Treponema may infect the brain via branches of the trigeminal nerve.”
Now, you will see why this "single-spirochete-becomes-multiple-strains" business is important later in this chapter, below, as regards Alan Barbour and Bioweapons, and how spirochetes creating multiple variants and all the individual spirochetes doing their own thing, varying their surface antigens on their own comes into play as far as ruining a person's immune system. And a ruined immune system is the DAMAGE and is the ILLNESS and is the specific goal of a bioweapon:
"Methods of using antipersonnel agents undoubtedly wary so that no uniform pattern of employment or operation is evident [make sure it does not produce antibodies, so assess the HLAs in the population you intend to abuse like the defecting Russian scientists at NYMC have been doing, is the short version- KMD]. It is likely that agents will be used in combinations so that disease symptoms will confuse diagnosis and interfere with proper treatment. It is also probable that biological agents would be used in heavy concentrations to insure a high percentage of infection [or just use the OspA vaccine- KMD] in the target area. The use of such concentrations [or the multiple infections it causes, due to the immunosuppression like HIV, Lyme, or LYMErix as acquired immune deficiencies - KMD] could result in the breakdown of individual immunity because the large number of micro-organisms entering the body could overwhelm the natural body defenses [or just infect or inject people with an immune suppressor like OspA from a tick or a syringe, and the reverse will happen: people will acquire multiple infections because their immunity is trashed by fungal OspA- KMD].
Do you see the disease now? ▲▲▲ ?? It's fungal (shed borrelial antigens are TLR2/1-agonists or fungal). It is about ovewhelming the immune system; it is about not producing identifiable antibodies; your bioweapons should be like a Trojan Horse, setting off other latent infections; your immune system now sucks (overwhelmed means turned off); you don't have "biofilms" at least of borrelia; Lyme was the "perfect stealth disabler;"... and then all of the CDC's ridiculous lies... From the CDC's lies alone, you could call this an accidental release of a bioweapon. But we have other circumstantial scientific evidence, as you will see...
Moron LLMDs Alert on “Biofilms”:
Use “Borrelia Staining” or “Borrelia Silver Staining” as search terms in PubMed and discover that Borrelia in vivo do not cluster at all, much less under a “Biofilm.”
Here is one. Look closely now, for the “clustered spirochetes hiding under a biofilm” (there is no such thing):
Demonstration of Spirochaetes in Patients with Lyme disease with a Modified Silver Stain
Here is another one by Paul Duray (same guy who revealed that congenital Lyme brain damage kills babies and the same guy who revealed to us that Lyme and LYMErix diseases cause a leukemia like illness and that the cells in the CSF of Lyme patients "look like Epstein-Barr transformed (mutated, pre-cancerous) cells:
PAUL DURAY >>> "Morphology of Borrelia burgdorferi: structural patterns of cultured borreliae in relation to staining methods.
"The microscopic recognition of Borrelia burgdorferi in biologic fluids and tissues is difficult and challenging because of low numbers of organisms occurring as single isolated spirochetes, the apparent lack of colony formation in tissues, and differing lengths and structural morphologies."
Additionally, biofilms are covered in TLR2/1 agonists so the body does not even see them at all any more, if they are there, in this post-sepsis disease called Chronic Lyme, with the multiple reactivated herpes viruses, etc and the expansion of tolerance to other toll-like-receptor-managed antigen types.
Okay, so REVIEW: Biofilms are NOT responsible for the persistent symptoms in Chronic Lyme Disease. Spirochetes, while permanent - no one sane argues that they're not, since they always were known to be incurable infections, even with arsenic - (RICOCHRON.htm). So, what is responsible?
Go to POST_LYME_SEPSIS_2014_SUMMER.htm and see the following, among others, FROM THE NIH!!!!
10) This is the NIH (NINDS’s MS-Lyme Group) group that discovered that *** OspA *** was the cause of the MS/New Great Imitator outcome of Lyme reporting in the New York Times in the summer of 2013 (Martin and Marques, 2006):
When Lyme Disease Lasts and Lasts – Jane Brody
"Complicating the picture is the fact that some people with PTLDS symptoms apparently never had Lyme disease in the first place, Dr. Marques said in an interview. There are other infectious organisms — Epstein-Barr virus, for example — that can produce similar symptoms and may be the real culprits."
Here are the NIH's 2 reports that say OspA (TLR2-agonist) is the cause of the MS/CFIDS/EBV-reactivated kind of Lyme (that also causes humoral immunosuppression),... and that as a result of exposure to OspA-like antigens (shed constantly in a process called blebbing, as revealed by CDC officer Alan "Stealth Bomber" Barbour), you might not even have anti-flagellar antibodies (TLR5-agonists):
Borrelia burgdorferi Induces TLR1 and TLR2 in human microglia and peripheral blood monocytes but differentially regulates HLA-class II expression.
Borrelia burgdorferi lipoprotein-mediated TLR2 stimulation causes the down-regulation of TLR5 in human monocytes.
12) NCI and US Army Ft Detrick Pathologist Paul Duray on the CSF cells looking like "Epstein-Barr-like transformed cells" in IDSA's 1989 Reviews Supplement on Spirochetal Diseases:
Rev Infect Dis. 1989 Sep-Oct;11 Suppl 6:S1487-93.
Clinical pathologic correlations of Lyme disease.
"Immature B cells can also be seen in the spinal fluid. These cells can appear quite atypical- not unlike those of transformed or neoplastic lymphocytes." --
Full Text: http://www.actionlyme.org/IDSA_CLINIPATH_DURAY.htm
NEW, by the NIH: "Surviving Sepsis: Detection and Treatment Advances"
By Carolyn Beans for the National Institutes of Health | August 18, 2014 08:43am ET
"Preventing Secondary Infections
"Some people who survive sepsis can develop secondary infections days or even months later. A research team that included Richard Hotchkiss, Jonathan Green and Gregory Storch of Washington University School of Medicine in St. Louis suspected that this is because sepsis might cause lasting damage to the immune system. To test this hypothesis, the scientists compared viral activation in people with sepsis, other critically ill people and healthy individuals. The researchers looked for viruses like Epstein-Barr and herpes simplex that are often dormant in healthy people but can reactivate in those with suppressed immune systems. [Sepsis Has Long-Term Impact for Older Adults, Study Finds]"
13) Duray again in 1992, in Steve Schutzer's review of the 1992 Cold Spring Harbor Conference on Lyme:
"On occasion, these atypical-appearing large lymphocytes have been misinterpreted in biopsy by several laboratories as cells of a malignant lymphoma or leukemia. Bb antigens, then, may stimulate growth of immature lymphocytic suibsets in some target organs, as well as in the cerebrospinal fluid (Szyfelbein and Ross 1988). Usual bacterial infections do not produce such lymphocytic infiltrates in tissue. ****These immunoblastoid cells in Bb infections at times resemble those found in Epstein-Barr virus infections.**** Does Bb reactivate latent virus infections in tissues? Do some tick inocula harbor simultaneous infectious agents (ixodid ticks can harbor Rickettsiae, Babesia microti, and Ehrlichia bacteria, in addition to Bb), producing multi-agent infections in some hosts? Further studies can clarify these issues by mans of tissue-based molecular probe analysis." -
Paul Duray, NCI, NIH, Ft. Detrick, at the 1992 Cold Spring Harbor Crooks' Conference, published in Steve Schutzer's Lyme Disease: Molecular and Immunologic Approaches. - book.
Yikes, I hope that is the end of THAT topic. God Save Us from stupid people who only will malpractice-treat rich people with long term antibiotics for post-sepsis immunosuppression, the reactivation of latent herpesviruses, and TL2/1 agonist (and expanded cross tolerance) tolerance, with their own little, “It must be babs, it must be bart, it must be today’s-flavor-of-whack-a-do coinfection number 467… and get a third mortgage, since the money from your second mortgage ran out, thanks, I love my new yacht….” abuse that you get from ILADS.org
BACK TO THE DNA SHELL GAME..... DO NOT SKIP THESE 2 PARAGRAPHS:
So, when talking about the DNA and RNA shell game played by the crooks - where the crooks use the wrong DNA to find “NO-LYME HERE!” and the correct DNA when they want to run to the patent office to claim a new species…- we have to go back to Chapter One of Cryme Disease (CRYMEDISEASE_CHP1.htm is the extension) which is about Taxonomy
The very first thing you need to know about Lyme is that it is technically called Relapsing Fever.
Here is why:
http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=138 <<< Click on the larger texted heading, "Borrelia" in the above link. You will see that the taxonomic categorization of these organisms is based on differences in their flagellin, because that is the genetic species distinguisher - and not OspA or any of the plasmid DNA. [All Borrelia are arthropod-borne. That is why they are called borrelia. That is why they're distinct from other spirochetes. And if they are parasites, they cause disease in some way.]
One must understand how these spirochetes are related,… and the phylogeny,… and in the end “they’re all just Borreliae and you really can’t even talk about species ,” Oscar Felsenfeld… But if you must, use the flagellar genes, since those is the species distinguishers, taxonomically.
On second thought... I have decided to add the links and the graphics to Phylogeny Basics so you don’t have to bother going to the Taxonomy database.
Here we see as the key report from the NIH's NLM's Taxonomy (Fukunaga, et al) database showing Burgdorferi is closest to anserina, an African bird borreliosis. They just happen to do this kind of African-Diseases-With-North-American-Vectors-kind of "Research" on Plum Island (verifiably):
Here we see the NIH Rocky Mountain Bioweapons Lab using anserina as an out-group, when in fact, it is the origin of the Plum Island burgdorferi group - African bird borreliosis - experimentally introduced to the microscopic bioweapony Ixodes, hard bodied tick:
More on Evolution and Expansion of the anserina-come-burgdorferi-Plum-Island phenomenon:
SUNY-SB on Lyme/Plum Island as the original outbreak area:
Evolution of a focus of Lyme disease
UPenn on Lyme being evolutionarily unlikely:
UNCOORDINATED PHYLOGEOGRAPHY OF BORRELIA BURGDORFERI AND ITS TICK VECTOR, IXODES SCAPULARIS:
“Despite the intimate association of B. burgdorferi and I. scapularis, the population structure, evolutionary history, and historical biogeography of the pathogen are all contrary to its arthropod vector.”
Durland Fish and his experiments with African diseases and vectors on Plum Island:
African Swine Fever Virus Infection in the Argasid Host, Ornithodoros porcinus porcinus
“Chiredzi/83/1 (Ch1) was isolated from Ornithodoros spp. ticks collected near Chiredzi, Zimbabwe (26), and was obtained from the Plum Island Animal Disease Center reference collection. Pretoriuskop/96/4/1 (Pr4) and Crocodile/96/1 (Cr1) were isolated from O. porcinus porcinus ticks collected from warthog burrows in Kruger National Park, Republic of South Africa, in September 1996. Nooitverwacht/96/6 (No6) was isolated from O. porcinus porcinus ticks collected from a warthog burrow in the Northern Province, Republic of South Africa, in September 1996.
CDC Lying about the viability of the cyst or spheroplast form of spirochetes,... and CDC lying about mycoplasma not being involved in Chronic Fatigue Syndrome:
CDC: "How to Dessicate and Weaponize your Borrelia :) "
CDC Throwing out the red blood cells (throwing out whole cells of any kind, including immune cells or white cells where the mycoplasma actually lived and used to be called epERYTHROzoa for that very reason:
"Absence of Mycoplasma species DNA in chronic fatigue syndrome"
"Plasma, the liquid portion of peripheral blood that is devoid of cells, is known to contain remnants of numerous physiological and disease processes. We used plasma DNA to detect and characterize bacterial 16S rDNA sequences in a group of individuals with CFS and a group of non-fatigued controls (Vernon et al., 2002). Whilst a variety of bacterial sequences were detected in both fatigued and non-fatigued groups, no Mycoplasma sp. 16S rDNA sequences were found."
So, that's all ▲ obviously a lot of research fraud performed by CDC staff already, and next, in the DNA Shell game you will see it is almost entirely CDC officers committing this fraud.
========== Now on to where the crooks play the shell game with the DNA and RNA ========
ALAN BARBOUR playing the DNA/RNA shell game...
You will want to look at The Patents chapter of Cryme Disease (http://www.actionlyme.org/CENTRAL_LYME_RICO_PATENTS.htm ) to see that CDC officer and former head of the NIH’s Rocky Mountain Bioweapons Lab (where there are tons of Relapsing Fever but no “Lyme”) Alan Barbour reported that basically “antigenic variation in one spirochete, times all the spirochetes you have leaves the immune system ‘overwhelmed’ with “an infinite number of new antigens,” which is a bioweapons technique well described by the US Army when speaking to Congress and can be seen on the Bioweapons pages of ActionLyme.org (http://www.actionlyme.org/120702.htm and http://www.actionlyme.org/BIOWEAPONS_ATTRIBUTES.htm )
ALAN BARBOUR states clearly that OspA undergoes true antigenic variation and that you cannot use this as a vaccine, and certainly not for DNA diagnostics as Klempner did in his “BREAKING NEWS!!!” Bogus re-treatment "study" that is now the data used by IDSA for their "Guidelines" from the summer of 2001.
"Antibody-resistant mutants of Borrelia burgdorferi: in vitro selection and characterization."
Says Barbour above: “Second, previous studies had shown antigenic differences in outer membrane proteins, OspA and OspB, between strains (21-26) and also true antigenic variation of these proteins within a strain (25, 27-30).”
That’s PubMed ID http://www.ncbi.nlm.nih.gov/pubmed/1339462
“Mutants” is code language. They’re all mutants. Antigenic Variation is the nature of the relapse in Relapsing Fever, so to call them mutants is silly and redundant, and not the least bit correct as you will see in Barbour’s patents and in the older data re the Single Spirochete outcome.
Here is what Barbour says in one of his patents about antigenic variation and "overwhelming the immune system":
"2.1 Methods of Treatment
"An important aspect of the invention is the recognition that Borrelia VMP-like sequences recombine at the vls site, with the result that antigenic variation is virtually limitless. Multiclonal populations therefore can exist in an infected patient so that immunological defenses are severely tested if not totally overwhelmed."
So you can’t use OspA for a vaccine, for post-treatment DNA diagnostics, or for Lyme case detection in antibodies. The only thing you can do or say about OspA is remember it means little except that it helped the normally non-borreliae-bearing Ixodes (hard bodied) ticks acquire a ligand (OspA-B plasmid) with which to attach to and invade the hard bodies of hard bodied (Ixodes) ticks. So, Lyme spirochetes were adapted probably on Plum Island to local vectors. Genetically, the Lyme spirochete is closest to anserina, an African bird borreliosis. All the more better to spread it around, to use a bird strain (see http://www.actionlyme.org/PIIB.htm and http://www.actionlyme.org/TRAINER_2012SUMMER.htm )
BARBOUR'S 1995 PATENT FOR SPECIFIC 16S RNA and Flagellin that GARY WORMSER DID NOT USE, WHILE BARBOUR SAYS THIS IS IN LONE STAR TICKS IN MISSOURI (that study is below in this chapter):
MARK KLEMPNER, playing the DNA/RNA shell game...
You have previously seen that the OspA gene undergoes antigenic variation (Alan Barbour, owner of the ImmuLyme vaccine OspA patent, above), is not found in all Borreliae and you can't use this DNA for anything, especially not vaccines or detection.
We move on to the Klempner "Study" which resulted in the “Guidelines” and where he references which DNA he used to assess "NO LYME IN LYME VICTIMS" (he doesn’t actually and the peer reviewers never noticed he did not list his primers):
So, what was that mysterious REFERENCE 21 above ^^ DNA that Klempner failed to report and the so called peer-reviewers did not notice?
So, the crooks – including Klempner in his bogus non-retreatment report that is now the basis of the IDSA “Guidelines” - say if the OspA gene is not there, there is no Lyme, right? Despite the fact that Lyme is a Relapsing Fever borrelia and OspA undergoes antigenic variation and not likely to be in the same form or produced by the exact same genetic code as one produced inside a tick, late in the disease in humans.
And here is Durland Fish using the correct primers to look for new species of Borreliae to patent in 2001:
A relapsing fever group spirochete transmitted by Ixodes scapularis... - PubMed – NCBI
Vector Borne Zoonotic Dis. 2001 Spring;1(1):21-34. Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. NCBI.NLM.NIH.GOV|BY SCOLES GA , ET AL.
"A 1,347-bp portion of 16S rDNA was amplified from a pool of infected nymphs, sequenced, and compared with the homologous fragment from 26 other species of Borrelia. The analysis showed 4.6% pairwise difference from B. burgdorferi, with the closest relative being Borrelia miyamotoi (99.3% similarity) reported from Ixodes persulcatus in Japan. Phylogenetic analysis showed the unknown Borrelia to cluster with relapsing fever group spirochetes rather than with Lyme disease spirochetes. A 764-bp fragment of the flagellin gene was also compared with the homologous fragment from 24 other Borrelia species. The flagellin sequence of B. burgdorferi was 19.5% different from the unknown Borrelia and showed 98.6% similarity with B. miyamotoi."
Yet, here we see a year later, Durland Fish using the WRONG DNA (OspA gene again), to assess treated mice, to determine if there is any Borrelia, coming to the conclusion that there is pretty much no Borrelia:
Detection of Attenuated, Noninfectious Spirochetes in Borrelia...
"PCR of DNA.DNA was isolated from individual ethanol-fixed nymphs or pooled larvae by means of the Isoquick DNA isolation kit (ORCA Research) and was resuspended in 20 μL of double-distilled H2O. Primers used for amplification were as follows: *** ospA *** (GenBank accession no. M57248, product amplicon coordinates 80–781): forward, 5′-AAAACAGCGTTTCAGTAGATTTGCCTGGTG-3′, and reverse, 5′-CAACTGCTGACCCCTCTAATTTGGTGCC-3′; BBE21.1 (GenBank accession no. AE000785, product amplicon coordinates 14663–14921): forward, 5′-AGAATTATGTCGGTGGCGTTGT-3′, and reverse, 5′-ATTAAAGCCGCCTTTTCCTTGGT-3′; and p37-47 (GenBank accession no. AE000794, product amplicon coordinates 1309–1457): forward, 5′-TTCTGATGGCACTGAGCAAACCA-3′, and reverse, 5′-AACCCTTTACACTTTCTTCGATTGCGCT-3′. The primer set for p37–47 has 100% homology to sequences in both B. burgdorferi strains B31 and N40, and the gene has been localized to lp28-1 in both strains [26, 27]. The primer set for BBE21.1 amplifies a unique region in lp25 of B. burgdorferi strain B31 downstream of BBE21 (amplicon coordinates 13403–14530) . BBE21 is located on a similar-size plasmid within B. burgdorferi strain N40 . We have been able to amplify by PCR the region corresponding to GenBank accession number AE000785, product amplicon coordinates 14195–14921, indicating that BBE21 and BBE21.1 reside on the same plasmid in N40 (authors' unpublished data)"
^^^ Those are plasmids, those "lp... " things. Plasmids are from where the variable surface protein antigens vary themselves. So, that is a classical Durland Fish type "bogus article."
GARY WORMSER playing the DNA/RNA shell game....
Next we are going to look at Gary Wormser who is in 1992 using the correct primers:
Diagnosis of early Lyme disease by polymerase chain reaction amplification and culture of skin biopsies from erythema migrans lesions.
Here Gary Wormser in 1999 when using the CORRECT PRIMERS (16S) finds most people are infected with more than one species of Borrelia and that you can't really use Barbour-Kelly-Stoener culture media (the only one anyone sells in the USA):
Genetic diversity of Borrelia burgdorferi in lyme disease patients ... - PubMed – NCBI
J Clin Microbiol. 1999 Mar;37(3):565-9. Comparative Study; Research Support, U.S. Gov't, P.H.S.
NCBI.NLM.NIH.GOV|BY LIVERIS D , ET AL.
And yet here we have Gary Wormser not finding Lonestari in Missouri because *** he is using the wrong primers*** [he is not using Barbour's lonestari patent primers (1995), and his results conflict with Telford's, below, in 2001 report showing Missouri Relapsing Fever is closest to theileri a cow relapsing fever, and Wormser is using a general 16S primer not specific to Borreliae species at all]; Wormser is also using an OspA gene primer which Barbour determined in 1995 was not in his new theileri-like (cow relapsing fever) patented Borrelia:
2005: Microbiologic evaluation of patients from Missouri with erythema migrans.
“PCR amplifications were performed in a 50-μL reaction mixture containing 10 mmol/L Tris-HCl (pH 8.3); 1.5 mmol/L MgCl2; 50 mmol/L KCl, 0.1% (w/v) gelatin; 100 μmol/L each of dATP, dGTP, dCTP, and TTP; 1.25 units Taq polymerase; and 20 pmol of each primer. Detection of borrelial DNA in patient specimens and ticks was accomplished by the nested PCR amplification of flaB using primers FlaLL, FlaLS, FlaRL, and FlaRS as described by Barbour et al . PCR of 16S rDNA was performed with broad-range eubacterial primers 8FPL and 1492RPL , which yields a product of ∼1.5 kbp. In cases in which no detectable product was obtained, second-round heminested PCR was performed with 8FPL and a reverse primer (519R: 5′-TTACCGCGGCTGCTGGC-3′) targeted at residues 535–518 (numbering corresponds to residues in the 16S RNA sequence of Escherichia coli) in 16S rDNA; this resulted in a fragment of ∼500 bp. Some specimens were also tested by PCR targeted at ospA (forward primer, 5′-CTGCAGCTTGGAATTCAGGCACTTC-3′; reverse primer, 5′-GTTTTGTAATTTCAACTGCTGACCCCTC-3′) and/or recA .”
WORMSER did not use the correct^^^ Borrelia-specific flagellin, or Borrelia-specific spacer genes as shown below by Telford and Barbour in his lonestari-like patent, and he did not use genus specific 16S RNA genes ("broad-range eubacteria"?), and he used a reverse primers for E coli ?? LOL.
SAM TELFORD's 2001 REPORT SAYING “Southern Lyme” is CLOSEST TO THEILERI OR BOVINE RELAPSING FEVER:
You can see that the ^^^ crooks have already sequenced the 4 similar strains of flagellar and genus specific 16S RNA spacer genes that are derived from a cow or bovine relapsing fever.
TO THIS DATE – from 1995 to 2015 - still, NO ONE IS USING BARBOUR'S RECOMBINANT LONESTARI FLAGELLIN OR 16S RNA GENE or any other of these gallblammed proper DNAs or RNAs TO DETECT HUMAN ILLNESS, NATURALLY.
They all know the only way to detect Lyme/Relapsing Fever is with specific recombinant flagellins from all the Borreliae, similar to Yale’s Lyme specific flagellin patented method, US 5,618,533.
ROBERT SCHOEN - 1995
J Immunol. 1995 Dec 15;155(12):5700-4.
An ospA frame shift, identified from DNA in Lyme arthritis synovial fluid, results in an outer surface protein A that does not bind protective antibodies.
Fikrig E1, Liu B, Fu LL, Das S, Smallwood JI, Flavell RA, Persing DH, Schoen RT, Barthold SW, Malawista SE.
Oh, you mean Lyme is a Relapsing Fever organism, so you can’t use the OspA gene for human treatment outcomes assessment, Huh Mr. Schoen, or to detect “Lyme” in EM rashes?
Here, next Yale's Robert Schoen (who says Lyme is not a real disease and needs no treatment) using 23S RNA primers to assure his RICO monopoly strain (and later patent with Dave Persing) is related to burgdorferi,...
Borrelia burgdorferi enzyme-linked immunosorbent assay for discrimination of OspA vaccination from spirochete infection.
and also reveals there is "Lyme" in the Southern and Western states in 1996.
Another report of this type is "Borrelia burgdorferi infection of the brain: Characterization of the organism and immune sera in the mouse model"
"The plasmid content of N40Br was different from that of the infecting strain implying either a highly selective process during infection or DNA rearrangement in the organism in vivo. "