Lyme disease tests should be performed every time a tick bite has occurred. You have to wait up to several weeks for the diagnosis to begin. Before that, it is possible to test the tick that has bitten us to see if it is the carrier of any disease. What is Lyme disease testing like and what is the cost of the tests?

Lyme disease tests(tick-borne bacterial disease caused by Borrelia burgdorferi and its varieties) should be performed as early as possible after the tick bite.

In the diagnosis of Lyme disease, specific diagnostic schemes are used, which are mainly based on serological methods.

However, taking into account the complex pathomechanism of the disease and the limitations of laboratory methods,Lyme disease can cause many diagnostic difficulties.

Therefore, there are more and more laboratory tests on the market that could solve this problem. Unfortunately, most of them do not have a proven diagnostic value and should not be used in the routine diagnosis of Lyme disease.

Contents:

    1. Early local Lyme disease - laboratory tests
    2. Early disseminated Lyme disease - Lyme test
    3. Late Lyme disease - what tests to detect Lyme disease?
    4. Lyme borreliosis - treatment monitoring
    5. Lyme disease - circulating immune complexes
    6. Lyme disease - lymphocyte transformation test (LTT)
    7. Lyme disease - CD57 test
    8. Berlioz - ELFA and CLIA test
    9. Lyme disease - examination of a tick removed from the skin

Early local Lyme disease - laboratory tests

At this stage, after contact with an infected tick, local infection occurs and the so-called wandering erythema (Latin erythema migrans), which is a symptom characteristic of Lyme disease.

The presence of erythema at the site of the tick bite is sufficient to diagnose Borrelia infection . The early local Lyme disease stage lasts from a few days to about 4 weeks, and the patient may develop flu-like symptoms.

It is the earliest stage of infection in which serological methods (ELISA and Western blots) testing the presence ofIgM and IgG antibodies may give false negative results. At this stage, the so-called serological window, i.e. the initial period of infection, in which the body does not yet produce antibodies against Borrelia.

The first IgM antibodies appear around 2 weeks of illness , but most people only detect their presence after a few weeks. In the case of a negative serological result, the test should be repeated approximately 3-4 weeks after infection.

ELISA and Western blot tests are reimbursed by the National He alth Fund, but the referring physician must have grounds for their order.Privately, the ELISA test costs from 30 to 100 PLN, and the Western blot from 80 to 200 PLN.

At a very early stage of infection, molecular PCR testing may be considered. The method detects bacterial genetic material (DNA), which is the most abundant in body fluids at the early stage of infection. The longer the time from infection, the lower the probability of detecting bacterial DNA.

However, it should be borne in mind thatin Poland there is still no standardization of molecular methods in the diagnosis of Lyme diseaseand therefore this method is not recommended as a diagnostic tool. Bmolecular supply is not reimbursed by the National He alth Fund and costs between PLN 160-200.

Worth knowing

Stages of Borrelia burgdorferi infection:

  • early local Lyme disease
  • early disseminated Lyme disease
  • late borreliosis

Early disseminated Lyme disease - Lyme test

In the early stage of disseminated Lyme disease, many organs, joints, muscles, nervous system and cardiovascular system are infected. Early Lyme disease develops with symptoms such as headaches, inflammation of the facial nerve, fatigue, or neck stiffness. This phase lasts for about a few weeks from the moment of infection.

In the early stage of disseminated Lyme disease, IgM antibodies are present. In the 6-8 weeks after infection, IgG antibodies also begin to appear. Therefore, the basis of diagnosis at this stage of infection are serological methods.

Serological diagnosis of Lyme disease begins with the screening stage, i.e. a sensitive ELISA test.

If the result is negative, it means no Borrelia spirochetes have been detected or the result is a false negative. When the result is positive or weak positive, confirmatory Western blot , which is highly specific, should be performed to confirm the results with the ELISA method.

In case of suspicion of neuroborreliosis, can be donemolecular PCR test from the cerebrospinal fluid. The PCR method is very sensitive, and considering that the number of Borrelia spirochetes in the cerebrospinal fluid is small, it may be helpful in making a diagnosis.

By itself, however, without prior serological testing, it does not constitute an infection diagnosis.

Late Lyme disease - what tests to detect Lyme disease?

The last stage of infection is late Lyme disease, in which there is a further spread of bacteria to organs and the formation of more and more serious complications of the infection. Late Lyme disease, joint Lyme disease or atrophic dermatitis appear. Late Lyme disease can last up to several years.

At this stage of the infection, only IgG antibodies are detected by serological methods.High levels of IgG against Borrelia spirochetes and the combination of symptoms indicate Lyme disease.However, in some cases (especially in people who received antibiotic therapy very quickly at an early stage of infection), IgM antibodies may persist even for several years. In such situations, these antibodies are not indicative of active infection.

What is Lyme disease?

Lyme borreliosis - treatment monitoring

One of the biggest difficulties in the diagnosis of Lyme disease is the lack of sensitive and specific laboratory markers that can assess the effectiveness of antibiotic therapy. The standard method to assess its effectiveness is the observation of disease symptoms.

Research indicates that the determination of the level of IgM and IgG antibodies against the spirochete VLsE / C6 antigen may be used to monitor the effectiveness of Borrelia infection.

Spiral protein VlsE / C6 causes a very strong and rapid response of the immune system. The addition of this antigen to the set of serological tests significantly increases the sensitivity of the test. The presence of IgG antibodies against the VLsE / C6 protein in patients with Lyme disease is:

  • 20-50%in patients at an early stage of infection
  • 70-90%in patients with early disseminated Lyme disease
  • almost 100%in patients with late Lyme disease

The test is not reimbursed by the National He alth Fund and costs PLN 80-150.

Lyme disease - circulating immune complexes

The basis for the diagnosis of Lyme disease is serological testing. However, there are clinical situations that may lead to a false-negative result. One of them is the presence of the so-called immune complexes that consist of Borrelia spirochetes and specific antigensIgM or IgG antibodies.

Immune complexes block the antibodies, making them impossible to detect by serological methods and therefore may cause false-negative results.This problem occurs mainly with very intense infections.

If a patient has obvious symptoms of Lyme disease, and the test results are negative, some laboratories offer a test that involves chemical breakdown of immune complexes.This procedure is aimed at releasing the antibodies and only then measuring their concentration.

Research indicates that the use of such a procedure prior to serological testing may significantly increase the detection of antibodies to Borrelia antigens. However, this method is not used routinely due to the lack of standardization in laboratories.

The price of the test ranges from PLN 110-210(for IgM or IgG antibody)and is not reimbursed by the National He alth Fund .

Lyme disease - lymphocyte transformation test (LTT)

The Lymphocyte Transformation Test (LTT) is used in various fields of medicine, including in transplantology, allergology and indirectly in the diagnosis of Borrelia spirochetes. In the diagnosis of Lyme disease, the test identifies the presence of immune cells (T lymphocytes) in the patient's blood that have been in contact with the bacteria. The test uses the ELISPOT technique, the principle of which is similar to the ELISA method, but is more sensitive than it.

Some people indicate that LTT testing for Borrelia spirochetes should be performed in order to:

  • confirmation of active infection in people with positive serological test results, but without symptoms clearly indicating the disease
  • confirmation of active infection in people with negative or borderline results in serological tests, but with clinical symptoms of the disease
  • monitoring the infection after antibiotic treatment
  • assessment of recurrence of Lyme disease

According to the recommendations of the Working Group published in 2014, both the LTT test and the determination of the level of antibodies in the circulating immune complexes are not recommended in the diagnosis of Lyme disease.

These methods still need to be validated (i.e. confirmed to be reliable) to be a useful diagnostic tool.

The price of the test is PLN 400-550 and it is not reimbursed by the National He alth Fund.

Lyme disease - CD57 test

The CD57 surface marker is a protein that occurs on the surface of immune cells - NK cells(Natural Killers).

Doctors from the ILADS group, i.e. the International Society for Lyme Disease and Related Diseases, believe that in the chronic phase of Lyme disease, the immune system is suppressed, which is reflected in a reduction in the population of NK cells with the CD57 marker.

They postulate that the determination of their number in the blood may be used to assess the activity of the disease. However, similar to the LTT method and the disruption of immune complexes, this method still requires validation.The price of the test is PLN 170 and it is not reimbursed by the National He alth Fund.

More about the CD57 test can be found here.

Berlioz - ELFA and CLIA test

In the diagnosis of Lyme disease, the ELISA(Enzyme-Linked Immunosorbent Assay) method is primarily used. However, many laboratories also use other serological methods such as ELFA (Enzyme Linked Fluorescent Assay) performed with Vidas® and CLIA (Chemiluminescent Immunoassay) performed with Liaison® analyzer.

What are the differences? The principle of operation of the three methods is very similar and uses specific antibodies and their ability to bind to antigens. ELFA and CLIA methods allow for the automation of analyzes and faster execution of tests in the laboratory. These methods are also able to detect lower concentrations of antibodies in the tested biological material.

When it comes to the credibility of the results obtained with three methods, they are almost the same.Positive or weak positive results with ELFA and CLIA methods, as for the ELISA method, should be confirmed by Western blot method.

Worth knowing

Lyme disease - examination of a tick removed from the skin

Some laboratories offer testing of a tick removed from the skin for the presence of Borrelia DNA. According to the recommendations of the Working Group published in 2014, such a study is absolutely not a diagnostic method of Lyme disease.

Although the risk of transmitting bacteria from ticks to humans is proportional to the time of sucking blood, the presence of spirochete DNA in the tick is not synonymous with infection, and its absence does not exclude infection.

Proponents of the method believe that it is helpful in making a decision about the introduction of antibiotic therapy in people with symptoms indicative of Lyme disease, but without the presence of migrating erythema.The price of such respondents ranges from PLN 100-500 and it is not reimbursed by the National He alth Fund.

References

  1. Laboratory diagnosis of tick-borne diseases. Recommendations of the Working Group: National ChamberLaboratory Diagnosticians, National Institute of Public He alth-National Institute of Hygiene, National Consultant in the field of infectious diseases, Department of Infectious Diseases and Neuroinfections, Medical University of Białystok, Polish Society of Virology, Warsaw 2014.
  2. Rychlik U. et al. Lymphocyte transformation test (LTT) laboratory diagnostics. Journal of Laboratory Diagnostics 2013, 49 (4); 425-428.
  3. Marques A. R. Laboratory Diagnosis of Lyme Disease - Advances and Challenges. Infect Dis Clin North Am. 2015; 29 (2): 295-307.
  4. Marques A. et al. Natural killer cell counts are not different between patients with post-Lyme disease syndrome and controls. Clin Vaccine Immunol. 2009; 16: 1249-1250.
  5. Krzemień P. J. The importance of the Vlse / C6 antigen as a marker in the diagnosis of early Lyme borreliosis and the study of its treatment effectiveness. He alth Problems of Civilization 2022; 11 (2): 87-92.
  6. Miąskiewicz K. et al. Proposition of a new methodological approach and interpretation of the results of antibodies against Borrelia burgdorferi - analysis of the antibody composition of circulating immune complexes. Rheumatology 2011; 49, 5: 328-334.
  7. Muraczyńska B. and Gałęziowska E. Controversies related to the diagnosis and treatment of Lyme disease in the world. General Medicine and He alth Sciences, 2015; 21 (4): 372-377.
About the authorKarolina Karabin, MD, PhD, molecular biologist, laboratory diagnostician, Cambridge Diagnostics Polska A biologist by profession, specializing in microbiology, and a laboratory diagnostician with over 10 years of experience in laboratory work. A graduate of the College of Molecular Medicine and a member of the Polish Society of Human Genetics. Head of research grants at the Laboratory of Molecular Diagnostics at the Department of Hematology, Oncology and Internal Diseases of the Medical University of Warsaw. She defended the title of doctor of medical sciences in the field of medical biology at the 1st Faculty of Medicine of the Medical University of Warsaw. Author of many scientific and popular science works in the field of laboratory diagnostics, molecular biology and nutrition. On a daily basis, as a specialist in the field of laboratory diagnostics, he runs the content department at Cambridge Diagnostics Polska and cooperates with a team of nutritionists at the CD Dietary Clinic. He shares his practical knowledge on diagnostics and diet therapy of diseases with specialists at conferences, training sessions, and in magazines and websites. She is particularly interested in the influence of modern lifestyle on molecular processes in the body.

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