The complexity of Lyme diagnosis
The Diagnosis of Lyme Disease is Complex and Requires Specialized Testing. In conventional healthcare, the diagnosis is based on symptoms combined with blood tests for antibodies against the bacteria. However, these commonly used tests in conventional medicine are only 34–59% sensitive, meaning many Lyme infections go undetected.
Clinical Diagnosis in Erythema Migrans
A characteristic symptom of Lyme disease is the red ring-shaped rash, known as erythema migrans. When this symptom is present, a clinical diagnosis is usually made, and further testing is not necessary. However, the diagnosis can be confirmed with PCR on a skin biopsy or a serological test.
Diagnosis in Chronic Skin and Joint Disorders
For patients with more chronic symptoms such as acrodermatitis chronica atrophicans (ACA) or Lyme arthritis, PCR combined with ELISA can be used to support the diagnosis. PCR helps provide direct evidence of the bacteria in skin or joint tissue. ELISA tests the antibody response of the immune system to the bacteria.
Challenges in Lyme Neuroborreliosis
Diagnosing Lyme neuroborreliosis, a neurological form of the disease, is particularly difficult. PCR and culture methods are too insensitive to be routinely used in clinical practice for this form of Lyme disease, as the bacteria are hard to isolate from nervous tissue in this way. Since early and accurate diagnosis is essential for treating Lyme neuroborreliosis, improved diagnostic tools are necessary.
Standard Serological Tests
The interpretation of serological results is complex. The relationship between antibody status and the actual infection is not always clear. The two most commonly used serological tests are enzyme-linked immunosorbent assays (ELISA or EIA) and immunoblots (IB). ELISA tests have higher accuracy (sensitivity) than immunoblots and are typically used first. Immunoblots are only used when ELISA is positive. This low sensitivity rate poses a significant challenge in conventional healthcare.
ELISA screens the blood for the total amount of IgM and IgG antibodies against the Borrelia bacteria. However, there are many variants of the test, and the quality varies significantly. IgM appears about three weeks after the onset of the infection, so testing immediately after a tick bite is not useful. The presence of IgM indicates an ‘acute infection.’ IgG typically appears weeks to months later. If you have a positive IgG result, it means you have previously been exposed to the Borrelia bacteria. It is estimated that around 50% of Lyme patients have negative ELISA results. Some Lyme patients may not produce (enough) antibodies because the bacteria are not always visible to the immune system or there was an issue during antigen presentation. Furthermore, ELISA only tests free antibodies, not antibodies already bound to Borrelia antigens. The test then comes back falsely negative, even though the patient is quite ill, because if the immune system does not produce sufficient antibodies, the Borrelia bacteria have free rein. Additionally, IgG continues to be produced even after the infection has been overcome. This test cannot accurately differentiate between a chronic infection and a resolved one.
IgM and IgG ImmunoBlot / Western Blot
These tests are used to determine if pathogen-specific antibodies are present in the patient's serum or plasma. Like ELISA, it is possible with this test to measure the reaction to specific Borrelia antigens, which improves specificity. It uses various bands that are specific to Borrelia (such as OspA/p31, OspB/p34). The different bands in the blot can provide additional information about the course of the disease, especially if the test is repeated at a later stage.
Diagnostic Innovations in Complementary Care
TickPlex is an innovative ELISA-based test that can detect persistent forms of Borrelia. The test contains an antigen specific to round bodies (persisters), a form in which the bacteria become invisible to the immune system. TickPlex Plus allows for the simultaneous determination of IgM and IgG antibodies for various bacterial and viral pathogens. TickPlex has a sensitivity of 95% and specificity of 98%, making it superior to traditional methods.
Borrelia EliSpot displays the actual activity of chronic and recent Borrelia burgdorferi infections by measuring T-cell response. The cellular immune response against Borrelia is characterized by a strong Th1 response (intracellular immunity), where Borrelia activates Th1-like cytokines such as interferon gamma (IFN-γ). This test is highly sensitive and can detect even one reactive T-cell. With a detection sensitivity of just one cell in 100,000, EliSpot is one of the most sensitive cellular tests available (20 to 200 times more sensitive than ELISA). EliSpot shows similar sensitivity to RT-PCR (Real Time PCR) analysis but detects the excreted protein instead of mRNA. EliSpot can be useful in monitoring therapies, as the test typically shows a negative result 4 to 8 weeks after completing effective therapy. EliSpot can also show the actual activity of various co-infections, such as Anaplasma, Bartonella, and Mycoplasma.
Lyme iSpot is also capable of detecting T-cell responses. The primary function of T-cells is to stimulate B-cells by producing specific and non-specific cytokines such as IFN-γ and IL-2. In general, IFN-γ is associated with an acute or active infection, while IL-2 is associated with immune memory and recovery. The relationship between the IFN-γ and IL-2 responses allows for a differentiated view of the immune status during and after infection.
Lymphocyte Transformation Test (LTT) measures the response of T-memory lymphocytes. Lymphocytes that have been in contact with Borrelia can only be measured within 45 days.
Natural Killer (NK) Cell Test (CD57+, CD56+, CD3-, CD19+): Several clinical and case studies have shown that chronic Lyme infections often coincide with changes in the cellular immune system. Evidence for this is a reduced number of NK cells (NK/CD3-, CD57+, CD56+). While acute Borrelia burgdorferi infections and other diseases have normal CD57 values, chronic Lyme patients often have fewer than 100 CD57 cells/µl. A low number of CD57 cells is mainly observed in patients where the nervous system has been affected, rather than those with tissue or skeletal complaints. The decline in CD57 cells persists until symptom improvements are achieved through antibiotics and/or other treatments. Conversely, a reduced CD57 is seen as a parameter for an active chronic Borrelia infection and a potential indicator that therapy has not yet been successful. CD19 (B-lymphocytes) specific cell surface antigens recognize and induce an antibody response. Low numbers of absolute CD19+ can indicate generalized B-cell lymphocytopenia, cellular immune deficiencies caused by stressful situations, corticosteroid therapies, chemotherapy, mycotoxins, etc.
Borrelia Extended Culture Testing (BEST) uses fluorescence techniques to test for the bacteria in blood cultures (reliable but very expensive). Unfortunately, Borrelia is not iron-dependent, prefers to dip into tissue, and is not always present in blood.
Nanotrap Test: Urine is examined for the presence of DNA particles from the Borrelia bacteria. The urine is processed so that the very low concentrations of DNA can be made visible.
Live Blood Analysis: Blood is directly examined under a dark field microscope for live bacteria. The Borrelia bacterium is easy to recognize due to its spiral shape, provided it is present in the blood.
Bioresonance: It is based on the premise that every cell/organ has a specific electromagnetic frequency. When there is disease or imbalance in the body, these frequencies may be disrupted. This disturbance can be measured with a bioresonance device. Microorganisms also have their own unique frequency, which can be measured and treated.
Phage Test: Bacteriophages are one of the simplest and most primitive forms of life—viruses. They are highly specific to their bacterial host. The presence of a Borrelia phage is 100% proof that Borrelia is present. Bacteriophages circulate in the blood, making the phage test suitable for patients in late stages when the bacteria are already hidden in the tissues.
Dutch Study
Because a positive two-tier serological test result is not evidence of active disease, a Dutch study (2) investigated the "usefulness" of the EliSpot. The study included 33 active/recent Lyme patients, 37 treated Lyme neuroborreliosis patients, 28 healthy individuals treated for an early manifestation of Lyme neuroborreliosis in the past, and 145 healthy individuals. The number of Borrelia-specific IFN-γ-secreting T-cells in active Lyme neuroborreliosis patients was compared with the number of Borrelia-specific IFN-γ-secreting T-cells in the other three groups. No significant difference was found in the number of B. burgdorferi B31-specific IFN-γ-secreting T-cells between the following three groups: active Lyme neuroborreliosis patients, treated Lyme neuroborreliosis patients, and those treated for an early manifestation of Lyme neuroborreliosis in the past. However, these three groups (ever infected with Borrelia) had higher numbers of B. burgdorferi B31-specific IFN-γ-secreting T-cells than untreated/never infected healthy individuals.
Conclusion of the study: EliSpot has no added value in diagnosing "Active Disease." This test cannot show a significant difference between active Lyme neuroborreliosis, treated patients, and healthy individuals who have been infected in the past.
What is striking is that this study does make a distinction between (ever) infected and never infected healthy individuals.
Isn’t this significant? Given the high number of people with severe symptoms, but a lack of recognition and treatment?
In my practice, I use the golden trio: EliSpot + CD57 + TickPlex.
A combination of EliSpot (current T-cell activity), CD57+ (NK-cell response), and TickPlex (humoral response, also for persistent forms) provides a broader insight into the immune response to potential pathogens. A comprehensive evaluation of both the antibody response and T-cell response to Borrelia infection can provide insights into the pathogenesis, treatment, and monitoring of the progress of Lyme disease.
Leeflang MM, Ang CW, Berkhout J, Bijlmer HA, Van Bortel W, Brandenburg AH, Van Burgel ND, Van Dam AP, Dessau RB, Fingerle V, Hovius JW, Jaulhac B, Meijer B, Van Pelt W, Schellekens JF, Spijker R, Stelma FF, Stanek G, Verduyn-Lunel F, Zeller H, Sprong H. The diagnostic accuracy of serological tests for Lyme borreliosis in Europe: a systematic review and meta-analysis. BMC Infect Dis. 2016 Mar 25;16:140. doi: 10.1186/s12879-016-1468-4. PMID: 27013465; PMCID: PMC4807538.
van Gorkom T, Sankatsing SUC, Voet W, Ismail DM, Muilwijk RH, Salomons M, Vlaminckx BJM, Bossink AWJ, Notermans DW, Bouwman JJM, Kremer K, Thijsen SFT. Een enzym-gekoppelde immunosorberende spottest die Borrelia burgdorferi B31-specifieke interferongamma-secreterende T-cellen kunnen actieve Lyme-neuroborreliose niet onderscheiden van Lyme Borreliosise uit het verleden: een toekomstgerichte studie in Nederland. J Clin Microbiol. 2018 26 mrt;56(4):e01695-17. doi: 10.1128/JCM.01695-17. PMID: 29367297; PMCID: PMC5869815.