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From the Townsend Letter
July 2017

How Finding a Lyme Spirochete in a Root-Canal Tooth Led to the Development of a New Test Panel
by Blanche D. Grube, DMD, and Leslie J. Douglas, PhD
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Lyme disease may have begun in a small New England village, but it is now a national, indeed global, phenomenon. In fact, Lyme has become the most commonly reported vector-borne disease in the United States. Thankfully, awareness of this problem is growing rapidly, both in the United States and abroad. This article intends to further advance this awareness. It will do so principally by discussing the detection of Lyme disease and its causative agents. At the end of our discussion, we also will consider some aids to successfully diagnose Lyme disease.
At this point, you may be asking yourself, "Why would a dentist, even a biological one, be discussing Lyme disease?" You would be right to ask such a question. After all, the mouth is not exactly a common place for a tick bite!
CoreBioticYet, our story starts right there: in the mouth. More specifically, it starts with a root canal.
It all began shortly after the passing of my very good friend and mentor Dr. Hal Huggins. He had developed what was called the "Full View Test," to identify the microorganisms present in the only part of the human body that cannot be effectively defended by the immune system: the mouth. He encouraged all dentists who had to remove unserviceable root canals to send them to Dental DNA for molecular-based testing. This was the laboratory that he established to identify microorganisms present in oral infections.
Shortly after Hal's death, I decided to change the lab's name to DNA Connexions. I did this because I believed it more accurately expressed the connection between oral health and the rest of the body. Well, after having sent an extracted root canal to the laboratory for testing, the lab manager excitedly called me with the news that the microorganism that causes Lyme disease had been detected. Subsequently, I asked our present lab manager at DNA Connexions, Dr. Leslie Douglas, to further develop our existing Lyme panel to include common tick-borne disease co-infectors. She did this with great zeal, and the result is the Lyme detection panel we currently offer. However, I am getting ahead of myself.

The Principal Lyme Infector and Associated Co-infectors
Before considering the detection and diagnosis of Lyme disease, it is necessary to first have a look at its principal causative agent. This will entail, furthermore, a brief consideration of associated co-infectors.
In the United States, the most common cause of Lyme disease is a bacterial spirochete called Borrelia burgdorferi. It is carried by two species of deer ticks (as opposed to wood ticks), Ixodes scapularis and Ixodes pacificus. It is the bite of these ticks that transmits B. burgdorferi to mammals, including humans.
However, B. burgdorferi can be associated with a multitude of tick-borne disease co-infectors. Our panel includes ten such organisms: Anaplasma phagocytohilium, Babesia divergens, Bartonella bacilliformis, Ehrlichia chaffeensis, Borrelia recurrentis, Borrelia miyamotoi, Babesia duncani, Bartonella henselae, Babesia microti, and Bartonella quintana.
I know, that reads like a tortuous tongue-twister! If only that were the extent of their malfeasance. Like B. burgdorferi, they all are bacteria, except for the Babesia co-infectors, which are parasites.

Different Available Testing Modalities
The Lyme issue is a great example of how science is all about distinctions!
It is one thing to have, or have had, a Lyme sphirochete in your system. In fact, B. burgorferi, the causative Lyme infector, can be a commensal organism. Essentially, that means that, provided its host stays healthy, it is perfectly content to "dine at the same table" (i.e., from the Latin words "cum" + "mensa") as everyone else, without causing any problems.
It is quite another thing to suffer from Lyme disease. An active Lyme infection requires a susceptible host, that is, one with an already compromised autonomic nervous system. What compromises the ANS? Stress, in all its forms, including mercury toxicity. And, who among us is not more or less adrenally overworked (i.e., "stressed")? And, how many of us still have mercury fillings?
So, there is the presence of a Lyme infector or co-infector(s), on the one hand, and the presence of an actual Lyme infection, on the other. These are two distinct realities. As a result, there also are two principal testing modalities available for Lyme detection, and they differ from one another in many respects.
Firstly, there is the standard serologically-based Western Blot test, which is blood-based, and which looks for evidence of an infection by examining antibodies produced in response to exposure. Unfortunately, it can be insensitive, both early in infection and following antibiotic or other therapies. So, this test panel looks for an immune response of the patient to an infection, a response which may be undetectable or even not there at all.
It was at one time believed that, in order to have an infection, one must have symptoms. However, Bartonella is an example of a microorganism that can cause a chronic bacteremia with few or no symptoms. Only a PCR test (as described below) would be able to tell whether the microorganisms are present or not. And, for the record, almost all ticks, fleas, and lice carry Bartonella. Another example of the old rules not being followed has to do with Babesia, which can cause a recurring infection long after the microorganisms are no longer detectable on a blood smear.
Secondly, there is the nucleic acid-based polymerase chain reaction (PCR) test. This can be either blood- or urine-based, and it detects the genetic footprint of the causative organisms. A PCR test is an in vitro nucleic acid (i.e., DNA) amplification process, obtained by means of thermocycling. Essentially, multiple genetically identical copies are produced from targeted genetic regions. The purpose of thermocycling is to make it easier to identify an eventual microorganism, or at least its residual DNA footprint.

Klaire LabsWhy We Prefer a Urine-Based Test Panel
A urine-based PCR panel has several principal advantages. The first is quite obvious. It is much easier to obtain urine from someone than it is to get their blood, especially in the case of children.
The second advantage concerns the nature of the urinary tract itself, which is a known site of spirochetal persistence. Essentially, that means they tend to reside there. This makes the urinary tract an ideal sampling reservoir.
Furthermore, a PCR test involves direct observation. We can only detect and report what is there. This means that it targets the genetic footprint of microorganisms, not the immune response of the patient in question. Here it is worth repeating: we can only detect, and therefore report, what is really in front of us.
Meanwhile, the Western Blot panel is an indirect testing method, looking for a person's immune response – which may or may not be there – rather than the presence of the causative organism itself.
Additionally, there is the fact that B. burgdorferi has evolved mechanisms to evade detection by the immune system. In other words, it hides.
Furthermore, B. burgdorferi is fairly individual specific. By that, I mean that everyone's reactionary response to the presence of this spirochete is unique. And, if no two of us react the same way, then it follows that each of us will respond just as uniquely to eventual treatment.
Also, B. burgdorferi remains in the blood stream only until the moment of dissemination. At that point, it moves from the blood to everywhere else in the body. This explains how it could have ended up inside a root-canaled tooth.
So, because the Western Blot test is based on blood, it could easily miss the positive presence of B. burgdorferi in other body tissues.

The DNA Connexions Answer
We encourage people to exercise, get a massage, or receive joint manipulation, and then to wait a full hour prior to providing their urine sample. When the joints and muscles are moved, the microorganisms begin to travel and ultimately wind up in the urinary tract. So, a urine-based test tends to offer the most comprehensive body sampling.
Patients sometimes ask us: "Why does my Western Blot read negative, while my PCR reads positive?" The answer is simple: they are two completely different tests. One is not necessarily better than the other, depending upon an individual's circumstances. They are just different. In other words, if you just want to test someone's blood, that is fine. However, if they are already immuno-compromised, then they may not be producing any antibodies for the Western Blot panel to detect as the targeted organism is not eliciting an immune response from its host. The bottom line is this: you are not likely to find what you are not seeking.

Lyme is a multi-tissue/multi-organ/multi-system disease. Moreover, it is multi-regional, having spread throughout the United States and abroad. Yet, only a small percentage of people who contract it ever see the typical bulls-eye rash that is associated with the infection. Nor do many even remember ever having been bitten by a tick.
However, even that having been said, we can offer suggestions for a practical harmonization of different test modalities. For example, in addition to serological testing, it may be beneficial for a given individual to begin with a PCR panel, especially one that includes a variety of co-infectors, to ensure the most comprehensive and targeted course of therapy. As I mentioned earlier, residual antibodies and not infections may be present in some cases.
Of course, even as a clinical diagnostic laboratory, DNA Connexions is not directly involved in either the medical diagnosis or treatment of Lyme disease. Instead, the laboratory offers its services as highly refined, molecular-based tools to aid medical professionals in making their own diagnoses on behalf of their own patients.

SUPPORTThe Townsend Letter is dedicated to examining and reporting on functional and integrative medicine. Our editorial content depends on support from readers like you, and we would appreciate your help to keep this content forthcoming. Please take this opportunity to contribute $50, or choose one of the other amounts listed on the next page, and ensure that our independent voices keep up the good fight against the skeptics, who would like to silence us and eliminate your medical freedoms.

Goodman JL, et al. Molecular Detection of Persistent Borrelia burgdorferi in the Urine of Patients with Active Lyme Disease. Infection and Immunity. January 1991; 269-278.
Jacomo V, et al. Natural History of Bartonella Infections (An Exception to Koch's Postulate).
Clinical and Diagnostic Laboratory Immunology. January 2002; 8-18.
Krause PJ, et al. Persistent and Relapsing Babesiosis in Immunocompromised Patients.
Clinical Infectious Diseases. 2008;46:370-6.
Lebech A-M, Hansen K. Detection of
Borrelia burgdorferi DNA in Urine Samples and Cerebrospinal Fluid Samples from Patients with Early and Late Lyme Neuroborreliosis by Polymerase Chain Reaction. Journal of Clinical Microbiology. July 1992; 1646-1653.

Blanche D Grube, graduated from Queens College, CUNY and received her doctorate from UMDNJ, now Rutgers School of Dental Medicine. She holds a second doctorate from Capital University of Integrative Medicine, Washington DC, and is a board-certified biological dentist and a past president of IABDM. Besides holding several fellowships, she is the owner and CEO of DNA Connexions, BIocomp Laboratories, Huggins Applied Healing and Centers for Healing.

Dr. Leslie J. Douglas completed her undergraduate studies in biology at the University of Hawaii at Hilo (UHH) before attending the University of Hawaii at Manoa (UHM), Department of Genetics and Molecular Biology. Currently, she is the Principal Investigator and Laboratory Manager of DNA Connexions, a Colorado-based company focusing on bacterial, viral, fungal, and parasitic molecular-based detection assays. Her main focus is the research and development of a PCR-based Lyme test inclusive of Borrelia burgdorferi and a number of prevalent tick-borne disease co-infections, as well as the ongoing development of various molecular-based assays. Dr. Douglas's research and patient demographics is yielding invaluable data to better understand the relationships between Lyme and other chronic conditions.

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