Borreliatesteistä ja niiden luotettavuudesta esiintyy erilaisia näkemyksiä.
Artikkelin lopussa on esitetty Saksassa esim. BCA klinikalla käytössä olevia borreliatestejä. Monien kohdalla Suomessa suoritetut borreliatestit ovat olleet negatiiviset ja esim. Saksan testit positiiviset. Allalolevan artikkelin mukaan Suomen testit ovat hyvin kehittyneitä. http://www.mtv3.fi/uutiset/kotimaa.shtm ... linikoilla
".....Saksassa joillakin yksityisklinikoilla käytössä olevista testeistä. ...nimenomaan nämä testit ovat huonoja, koska ne antavat vääriä positiivisia tuloksia".
Useiden näkemysten mukaan kuitenkin nimenomaan perinteiset vasta-ainetestit ovat taudin kaikissa vaiheissa varsin epäluotettavia ja siksi tarvitaan uudenlaisia testejä. Vuonna 2011, FDA; Amerikan Elintarvike- ja Lääkevirasto ja CDC; Yhdysvaltojen tartuntatautien valvonta- ja ehkäisykeskus, hyväksyivät ELISPOT LTT tekniikan käytön. tuberkuloosiin. Nyt tätä tekniikkaa käytetään useiden eri taudinaiheuttajien tutkimiseen.
Tekniikka ei ole käytössä Suomessa Borrelioosin diagnostiikassa ./size]
Borrelia Elispot-LTT (LymphocyteTransformationsTest)
The Elispot-LTT method has been approved by the FDA in May 2011 for M. tuberculosis (Not ITT or MELISA)
The FDA argues in this paper:
"... A positive result (in the Elispot-LTT) suggests that an infection is likely, a negative result that an infection is unlikely..."
"... Results (of the Elispot-LTT) can be available within 24 hours (ITT or MELISA not)..."
A Borrelia infection does not only activate the humoral immune response, but also activates T-lymphocytes at the same time. Once Borrelia bacteria are not active anymore, the T-cellular immune response is not present.
It is not possible to test the treatment success by Borrelia antibodies, because the 'titer" or antibodies can be measured in the blood over years. Furthermore, Lyme infections in Stage I (e.g. 'bulls-eye rash' or 'summer flu') only show antibodies in the blood after weeks and sometimes do not show them at all.
The Borrelia Elispot-LTT eliminates these problems. The test reflects the actual, current Borrelia burgdorferi activity of chronic and also acute Lyme infections. The Elispot-LTT is highly sensitive and can detect even one single Borrelia-reactive T-cell in the blood. The Elispot-LTT is very helpful when monitoring a chronic or acute Lyme therapy. The Elispot-LTT should usually become negative about 6 to 8 weeks after completion of an effective therapy.
Artkkeli LTT-tekniikastta eri mikrobeja tutkittaessa:
Elispot®-LTT: FDA and CDC approved LTT technique in U.S.
Actual T-cellular activity in the blood against Borrelia burgdorferi, Chlamydia pneumoniae, Chlamydia trachomatis, Ehrlichia/Anaplasma, Epstein-Barr-Virus, Yersinia
In May 2011 the U.S. Food and Drug Administration (FDA) and the Centers for Disease Control and Prevention (CDC) have approved the Elispot®-LTT (T-Spot) technique beneath the QuantiFERON® TB Gold In-Tube test. Both tests represent Interferon-Gamma Release Assays (IGRAs) in form of Lymphocyte Transformation Tests (LTT). No other laboratory T-cell tests have been approved (ie: MELISA® or ITT® techniques) in the field of all Lymphocyte Transformation Tests (LTT) by the FDA/CDC yet. In the paper of the CDC regarding Interferon-Gamma Release Assays (IGRAs) from May 2011 the CDC says: "... A positive result suggests that an infection is likely, a negative result that an infection is unlikely...", and "...results can be available within 24 hours..."
The Elispot®-LTT is available for the following infections:
- Borrelia burgdorferi
- Chlamydia pneumoniae
- Chlamydia trachomatis
- Yersinia species
LTT in Borrelia Testing
A Borrelia infection leads to a vitalisation of T-lymphocytes apart from the humoral immune answer. The T-cellular immune response vanishes as soon as the Lyme disease is not active anymore.
A therapy success control of a Lyme infection is not possible by the Borrelia antibodies, as the “titer” of antibodies in the blood can be found for years after an infection. Additionally in stage I (e.g. “bull´s eye rash” or “summer flue” after a tick bite) antibodies can be found after several weeks or in stage III they cannot be found in every case (weak immune system). These diagnostic gaps are closed by the Elispot-LTT for Borrelia, which detects the actual cellular activity against Borrelia burgdorferi in chronic and also acute Lyme disease. The Elispot is so sensitive, that even a single Borrelia can reactivate T-cells in the blood. The Elispot is 20- to 200-fold more sensitive than an ELISA-test on Borrelia and will already find 1 reactive T-cell in 100.000 lymphocytes. The Elispot for Borrelia is very important for controlling a therapy of a chronic or acute Lyme infection. In general the Elispot-LTT is going to be negative app. 6 to 8 weeks after the end of a successful therapy.
Advantages of the Elispot-LTT for Borrelia as performed by Infectolab in contrast to other lymphocyte transformation or proliferation tests from other laboratories are:
· The result is available within 5 days (Other similar tests: 2 – 3 weeks)
· The use of cell stabilising CPDA-tubes means a stability of 3 days for the measured cells after taking the blood (others use Heparin for a stability of only 24 hours)
· Better reliability than the different "unspecific" tests using the proliferation of T-cells (e.g. ITT®)
This offers a significant improvement of the stability of the T-cells and a very quick decision possibility for Lyme treating physicians to extend a therapy or to switch to a new treatment option for Lyme disease!
Elispot-LTT for Borrelia:
Material: 3 x 9 ml CPDA-tubes (yellow cap, kept at room temperature, do not cool or centrifuge)
Time required for analysis: 2 days (Results in 5 days)
Indications: - Diagnosis of chronic Lyme disease
- Diagnosis of acute Lyme disease
- Decision for the length of Lyme disease therapies
- Success control of therapies after Lyme treatment
http://www.aacc.org/events/Annual_Meeti ... 9-B150.pdf
Chenggang Jin, MD, PhD/Bradley Bush, ND
[size=150]Development and Validation of a Novel Interferon-γ ELISPOT Assay for Sensitive and Specific Detection of Antigen-Specific T cell Response to Borrelia burgdorferiThe enzyme-linked immunospot (ELISPOT) technology has proven to be extremely sensitive in detecting antigen specific reactive T cells and has been applied in laboratory diagnostic for Tuberculosis approved by FDA. A novel T-cell based assay for diagnosis of Lyme disease -Lyme ELISPOT was successfully developed and validated.
Learning Objectives: After the presentation, an individual will be able to:
⦁ Describe ELISPOT assay technology
⦁ Contrast ELISPOT assays to conventional antibody detection methods utilized for the diagnosis of Lyme disease
⦁ Evaluate the potential application of ELISPOT as a diagnostic tool for Lyme disease
Background: Lyme disease, caused by infection with the spirochete Borrelia burgdorferi, is an emerging infectious disease in the United States that has become an important public health problem. Both B-cell immunity and T-cell immunity develop in natural infection with Borrelia burgdorferi. Detection of specific antibody response mediated by B cells against Borrelia burgdorferi is utilized conventionally in aiding the clinical diagnosis of Lyme disease. However, the limitation of these antibody-based immunoassays is that they have low sensitivity and specificity, causing significant false negative and false positive results. Furthermore, Borrelia specific antibodies cannot be detected at the early stage of infection and in a fraction of seronegative Lyme patients who lack Borrelia specific antibody responses. In contrast, Borrelia specific T-cell based immune assays have not yet been well developed. Thus, highly sensitive and specific T-cell based clinical laboratory assays are needed to help in diagnosing Lyme disease in conjunction with antibody-based immunoassays. The enzyme-linked immunospot (ELISPOT) technology has proven to be extremely sensitive in detecting antigen specific reactive T cells and has been applied in laboratory diagnostic for Tuberculosis approved by FDA. We here explore the potential application of ELISPOT as diagnostic tool for Lyme disease.
Objective: The aim of this study is to develop and validate a novel T-cell based assay for diagnosis of Lyme disease using newly developed digitalized ELISPOT technology.
Methods: To develop the novel T-cell based diagnostic assay for Lyme disease, we detected the Borrelia antigen-specific memory T cells that were activated ex vivo by recombinant Borrelia specific antigens, using Th1 cytokine Interferon-γ ELISPOT at the single cell level. The human peripheral blood mononuclear cells (PBMC) were stimulated with single or a combination of recombinant Borrelia specific antigens, DbpA, OspC, p100 and VlsE. In addition, we added costimulatory cytokine IL-7 into the cell culture to increase the detection of T memory cells. The results of ELISPOT were analyzed using CTL S6 Ultimate-V Analyzer/BioSpot 5.0 Software and reported as IFN- γ Spot Forming Units (SFU). To validate the Lyme ELISPOT assay, a cohort of 21 clinically diagnosed Lyme patients and 45 healthy control subjects were further studied and compared with Western Blot test. The performance of the Lyme ELISPOT assay, including clinical sensitivity, clinical specificity, accuracy and precision, is also evaluated.
Results: The frequency of Borrelia specific T memory cells can be detected by Interferon- γ ELISPOT and therefore can be used as a biomarker for Borrelia infection. The detection of antigen specific T cells was significantly increased by a combination of recombinant Borrelia antigens and addition of constimulatory cytokine IL-7. The signal enhancing effect of IL-7 was observed even at saturating antigen concentration in terms of frequency, but IL-7 did not increase the amount of IFN- γ secreted by individual cells. A strong correlation was observed between ELISPOT and IFN- γ concentration measured by Bio-plex suspension system (R=0.8, P<0.0001). The Lyme ELISPOT assay cut-off value was determined using Receiver Operating Characteristic (ROC) curve analysis and it was found that a cut-off value of >25 PFU maximized assay sensitivity and specificity. It has a significantly higher specificity (96%) and sensitivity (76%) compared with Western blot (Sensitivity 24%). The results also demonstrated that there was dissociation between B cell response and T cell response during Borrelia infection, suggesting a comprehensive immunological diagnostic panel should include both B cell and T cell diagnostics. Further studies will include more Lyme patients, other related diseases and independent studies by other laboratories.
Conclusion: A novel T-cell based assay for diagnosis of Lyme disease -Lyme ELISPOT was successfully developed and validated. This newly developed Lyme ELISPOT assay may be a helpful laboratory diagnostic test for Lyme disease, especially for seronegative Lyme patients. A comprehensive evaluation of both antibody response and T cell response to Borrelia infection will provide new insights into the pathogenesis and diagnosis of Lyme disease.
Lymfosyyttitransformaatiotestistä/Borrelioosista lisää esim. http://www.cdc.gov/lyme/stats/chartstab ... yyear.html
Yound, J.D. Underreporting of Lyme disease. N Engl J Med. 1998. 338(22):1629-1629. http://www.cdc.gov/hiv/topics/surveillance/basic.htm http://www.cdc.gov/ncidod/dvbid/westnil ... tailed.htm http://www.who.int/influenza/human_anim ... 1cases.pdf
Maloney, E.L. The Need For Clinical Judgment in the Diagnosis and Treatment of Lyme Disease. Journal of American Physicians and Surgeons. 2009. 14(3):82-89.
Lehmann, P.V., Zhang, W. Unique Strengths of ELISPOT for T Cell Diagnostics. In: Alexander Kalyuzhny, E. Handbook of ELISPOT: Methods and Protocols, Methods in Molecular Biology, vol. 792. 2nd Ed. New York, NY: Spriner Science+Business Media, LLC; 2012: 3-23.
T-Spot.TB 96 [Package Insert]. Oxfordshire, UK: Oxford Immunotec Limited; 2009.
Tary-Lehmann M., Hamm, C.D., Lehmann, P.V. Validating reference samples for comparison in a regulated ELISPOT assay. In: Uma Orabhakar and Marian Kelley Eds. Validation of Cell-Based Assays in the GLP Setting: A Practical Guide. 1st Ed. West Sussex, England: John Wiley & Sons. Ltd; 2008: 127-146.
Forsberg, P., Ernerudh, J., Ekerfelt, C., et al. The outer surface of Lyme disease borrelia spirochetes stimulate T cells to secrete interferon-gamma (IFN-γ): diagnostic and pathogenic implications. Clin Exp Immunol. 1995; 101:453-460.
Ekerfelt C., Forsberg P., Svenvik, M., et al. Asymptomatic Borrelia-seropositive individuals display the same incidence of Borrelia-specific interferon-gamma (IFN-γ)-secreting cells in blood as patients with clinical Borrelia infection. Clin Exp Immunol. 1999. 115: 498-502.
Dattwyler, R.J., Volkman, D.J., Luft, B.J., et al. Seronegative Lyme Disease- Dissociation of Specific T- and B-Lymphocyte Responses to Borrelia burgdorferi. New England Journal of Med. 1988. 319(22): 1441-1446.
Dressler, F., Yoshinari, N.H., Steere, A.C. The T-Cell Proliferative Assay in the Diagnosis of Lyme Disease. Annals of Internal Medicine. 1991. 115:533-539.
Martinuzzi, E., Scotto, M., Enee, E., et al. Serum-free culture medium and IL-7 costimulation increase the sensitivity of ELISPOT detection. Journal of Immunol Meth. 2008. 333: 61-70.
Seriburi, V., Ndukwe, N., Chang, Z., et al. High frequency of false positive IgM immunoblots for Borrelia burgdorferi in clinical practice.Clin Microbiol Infect. 2012 Dec;18(12):1236-40.
Kuerten, S., Batoulis, H., Recks, M.S., et al. Resting of Cryopreserved PBMC Does Not Generally Benefit the Performance of Antigen-Specific T Cell ELISPOT Assays. Cells 2012, 1(3), 409-427
*Patent Pending (USPTO Application No: 61779064)
Lyme disease is an increasingly common condition that can have debilitating effects if not diagnosed and treated appropriately. A comprehensive approach to diagnosis will lead to the most positive outcomes and healthcare savings; however, testing options thus far have the potential for false negative results, making diagnosis difficult. NeuroSciences testing options (iSpot Lyme including Western Blot analysis) can aid in the diagnosis of B. burgdorferi infection and allow for early detection leading to earlier treatment. iSpot Lyme is a highly sensitive T cell-based enzyme-linked immunospot (ELISPOT) method which enumerates the B. burgdorferi-specific activated effector/memory T cells. This test represents a breakthrough in diagnostic accuracy, and can increase the speed of diagnosis and treatment leading to improved clinical outcomes.
-----------------------------------------------------http://www.mtv3.fi/uutiset/kotimaa.shtm ... linikoilla
Professori: Suomessa paremmat borrelioositestit kuin Saksan klinikoilla
Punkkitauti borrelioosista vuonna 1996 väitellyt professori kehuu Suomessa käytössä olevia borrelioositestejä.
Suomen testejä on kritisoitu julkisessa keskustelussa, ja ja ihmisten on jopa kerrottu matkustavan testeihin ulkomaille.
Turun yliopistollisen keskussairaalan infektiotautien ylilääkäri, infektiotautien professori Jarmo Oksi sanoo, että Suomessa käytössä olevat vasta-ainetestit borrelioosin diagnosoimiseksi ovat hyvin kehittyneitä.
– Väittäisin, että meillä käytetään todennäköisesti parempia testejä kuin suurimmassa osassa muuta Eurooppaa. Uskoisin, että juuri missään ei ole saatavilla parempia testejä, Oksi sanoo.
"Toinen testi antaa melkein kenelle tahansa positiivisen"
Oksi haluaa sanoa sanansa myös Saksassa joillakin yksityisklinikoilla käytössä olevista testeistä. Oksin mukaan nimenomaan nämä testit ovat huonoja, koska ne antavat vääriä positiivisia tuloksia.
– Joissakin Saksassa olevista yksityisklinikoista näytetään käytettävän niin sanottua lymfosyyttitransformaatiotestiä, joka on vähän eri asia kuin vasta-ainetesti ja selvästi ongelmallisempi. Tämä testi näyttää positiivista, jos on koskaan vähänkään törmännyt borreliabakteeriin, vaikkei bakteeria nykyään kehossa olisikaan, Oksi tyrmää.
Lymfosyytit kuuluvat ihmisen valkosoluihin, ja Oksin mukaan lymfosyytit voivat siis reagoida borreliabakteeriin, vaikka bakteeria ei ole elimistössä.
Oksi myöntää, että borrelioosin diagnosoiminen voi olla vaikeaa ja että Suomessa käytössä olevat vasta-ainetestitkään eivät ole täydellisiä.
– Ensimmäisen kahden kuukauden aikana punkin puremasta vasta-aineet eivät ole ehtineet vielä nousta. Toisaalta voi olla täysin terve, vaikka vasta-ainetesti olisi positiivinen. Vasta-aineet nousevat todennäköisesti kuitenkin hitaammin kuin lymfosyyttitunnistus, joka saattaa antaa melkein kenelle tahansa positiivisen tuloksen.
-----------------------------------------------------------------Saksan Borrelioosiklinikan testit:
http://www.b-c-a.de/fileadmin/img/bca/6 ... .07.11.pdf
Ihr kompetenter Laborpartnerhttp://www.infectolab.de
GmbH & Co. KG
Dr. med. Armin Schwarzbach
Facharzt für Labormedizin
, ein Geschäftsbereich der BCA-clinic Betriebs GmbH
& Co. KG
Geschäftsführer: Dr. med. Carsten Nicolaus, Dr. med
. Armin Schwarzbach
Amtsgericht Augsburg HRA 15697
Tel. +49/ 0821/ 455 074-0
Fax +49/ 0821/ 455 074-1http://www.infectolab.de
Kreissparkasse Augsburg (BLZ 720 501 01)
IBAN: DE48 7205 0101 0000 0198 85
Laboratory services and diagnosis for Lyme disease
- The detection of tick-borne infections makes high
demands on laboratory analysis in
combination with the diagnostic findings (anamnesis
It is important to detect Borrelia infections at an
early stage. The earlier the detection the simpler
the treatment measures (usually antibiotics) and th
e shorter the ordeal of the infected patients.
This is how you can diagnose Lyme Disease
a Lyme disease infection progresses in 3 stages dep
symptoms and ailments
(time specification after the tick bite):
(after days up to weeks): “bull’s eye rash“ (“Eryth
migrans“, only in 40-70% of all cases), Borrelia l
headache, fever, sweating,
“Summer flue“ (app. 20% of all
cases), exhaustion and fatigue, facial palsy (espec
(after weeks up to months): inflammation of the bra
spinal marrow; any nerve in the human body, inflamm
the joints (“arthritis”), joint and muscle pain, in
flammation of the
eye, liver and kidneys, myocarditis, pericarditis,
Borrelia burgdorferi – a
spirochete bacteria with a range
of over 300 proven species world
(after months up to years): Thinning of the skin at
the back of the hand, (“Acrodermatitis chronica
atrophicans“), Borrelia lymphocytoma (ear, nose, sc
rotum), lethargy, fatigue, paraesthesia, cognitive
dysfunction, muscle inflammations, joint inflammati
ons and swelling, tendon inflammations,
inflammation of the bursa, vasculitis, myocardinal
Symptoms of Lyme disease occur in thrusts with alte
rnating intensity and appearance in contrast
to a classic organic illness. Many patients also su
ffer from slightly higher temperature during those
thrusts. Co-infections with other bacteria and viru
ses have been increases over the past years and
often lead to a complicated course of the disease.
Often the tick bite is not detected early enough
or the acute treatment by the attending physician i
s not sufficient. The chronic Lyme disease
patients often go through a true “martyrdom” as the
y do not only suffer from actual physical and
mental ailments, but also from not receiving a reli
able diagnosis and the fact that their illness is n
There are three main reasons why a Lyme infection i
s not immediately detected in daily practice,
so that a treatment at an early stage is inhibited:
1. There is no
bull’s eye rash
(“Erythema chronicum migrans“)! Research has prov
this classic symptom of Lyme disease only appears i
n 40% to a maximum of 70% of all
No tick bite detected
! Such a bite can be induced even by very small tic
ks (larvae). Or it
was not detected because there was no specific skin
reaction. In addition, the scientific
community now assumes that Borrelia bacteria can al
so be transmitted by infected insects.
Only conventional blood tests
have been performed. This was either too early
(antibodies can be tested positive only after a per
iod of up to six weeks after the tick bite)
or there have been absolutely no antibody productio
n in the body or the cellular stage was
not or not sufficiently tested (necessary tests are
-LTT and CD3-/CD57+ cells).
The BCA-clinic Augsburg (
) is specialised in the diagnosis and therapy of ti
diseases. In case of a suspected tick-borne disease
the diagnosis is performed by experienced
physicians of the “Medical Partnership” cooperating
with the BCA-clinic. The diagnosis is based on
an extensive anamnesis in which precise ailments an
d the antecedent are recorded and reviewed
along with a physical examination (=
). Additionally, the physician will initiate
specific laboratory tests of your blood, which will
be undertaken and analysed in the specialised
laboratory of the BCA-clinic.
is crucial for the detection
of Borrelia. There is a difference between the test
s of the
NK-cells). Both levels have to be examined at the s
time when a Lyme disease or an apparent illness is
1. Laboratory testing of the humoral level (antibo-
Borrelia IgM- and IgG-EIA (Enzymimmuno-assay)
as well as Borrelia IgM- and IgG-Immunoblot
2. Laboratory-testing of the cellular level:
-LTT (Borrelia Elispot Lymphocyte Transformation
Test), CD3-/CD57+ cells (NK-cells)
Explanations of these tests:
1. Antibodies – humoral level: In case of the Borre
examinations up to 19% of the antibodies results in
the EIA are
falsely negative due to the minimal sensitivity (re
of the EIA in contrast to the Immunoblot. Therefore
essential to check the Borrelia IgM- and IgG-Immuno
with the Borrelia IgM- and IgG-EIA (even with a neg
Important: The laboratory has to always tests for V
major protein-like sequence Expressed) in EIA and
Immunoblot. VlsE describes the characteristics of
as a “chameleon“, which permanently changes the sur
structure VlsE in vivo to resist the detection via
system. VlsE has the highest sensibility for the an
Beware of positive antibodies constellations!
Armin Schwarzbach M.D.,
medical specialist for laboratory medicine,
head of the BCA laboratory and
specialist for Lyme and co-infections.
Borrelia IgM- as well as IgG-antibodies can remain
in the body for
months or years even without an active Lyme disease
Hence a positive Borrelia antibodies test does not
give any evidence
of the activity of a Lyme infection, but gives only
one conclusion: There must have been a tick bite o
bites in the past during which Borrelia bacteria ha
ve been transmitted – no more or less!
2. Cellular level:
provides information about the current activity of
the Borrelia bacteria and is
20 to 200 fold more sensitive than a EIA-antibodies
indicate the extent of immune suppression during a
chronic Lyme disease and are the
prognostic factor during and after the antibiotic t
The complete laboratory diagnosis is very complex a
nd - considering the co-infections - has to be
put together like a mosaic. This demands experience
d Lyme disease analysts who can diagnose
and evaluate all other infections as well.
Armin Schwarzbach, M.D., PhD, is head of laboratory
medicine and diagnostics in the BCA.-clinic.
Dr. Schwarzbach is very experienced in laboratory m
edicine and has been a specialist in Lyme
disease and co-infections for years.
The medical laboratory diagnosis is primarily orien
tated on the guidelines of the German Society
for Hygiene and Medical Microbiology (MiQ12 Lyme-Bo
rreliose). However, the complexity of a
Lyme disease infection is not sufficiently covered
by this and needs additional laboratory testing.
herefore further internationally accepted methods
are applied which are of considerable
importance for the patient before and after the the
rapy (e.g. for the cellular level: Elispot
the CD3-/CD57+ cells).
The laboratory integrated in the BCA for the analys
is of vital parameters ensures, especially in the
case of the medical treatment (e.g. infusion therap
y), that the physicians of the medical
partnership make further therapy decision as soon
as possible according to the blood results.
In practice, the following laboratory constellation
s indicate a Borrelia infection:
(1) Positive antibody detection and positive cellul
-LTT and/or CD57+) – often active infection
(2) Positive antibody detection without positive ce
llular test findings – What kind of indications
are shown by the clinical findings (symptoms)?
(3) Negative antibodies detection, but positive cel
lular test results
- Indication for an active Lyme disease at an early
stage, but also chronic stage – what
kind of indications are shown by the clinical findi
Attention: A negative antibody detection in the EI
A and/or Immunoblot does not give evidence of
a Lyme infection! Because: the antibodies productio
n of a Lyme disease in stage I needs several
weeks, 10 to 14 days at a minimum. Thus the immedia
te measurement of cellular activity in
-LTT is a compulsory necessity, as normally the cel
lular activity precedes the humoral
activity in stage I of a Lyme disease infection.
In practice the following combinations of blood tes
ts have proven useful:
Laboratory diagnosis stage I
(Costs: 393,44 €
plus extra charges.
1. Borrelia IgG- and IgM-EIA incl. VlsE
2. Borrelia IgG- and IgM-Immunoblot
3. Borrelia Elispot
Laboratory diagnosis stage II and
(Costs app.: 544 €
plus extra charges.
1. Borrelia IgG- and IgM-EIA incl. VlsE
2. Borrelia IgG- and IgM-Immunoblot
3. Borrelia Elispot
4. CD3-/CD57+ cells
“ (evaluation of progression) of Lyme disease durin
g and after an antibiotic or
holistic approach to therapy: The above mentioned p
arameters also have to be checked during
Recommended laboratory diagnosis during the therapy
Performance of above mentioned 3 tests
4 weeks after beginning of therapy and 8
weeks after the end of therapy.
Stage II and III
Performance of above mentioned 4 tests after
beginning of therapy every 8 weeks as well as
8 weeks after end of therapy.
Borrelia antibodies or titer cannot be “eliminated”
, but they can exist in the
blood for months or even years. After a successful
antibiotic therapy Elispot
-LTT as well as
CD57+ cells should have come to a “normal” level 8
weeks after the end of therapy. But
of a symptom free status after treatment there migh
t be still positive findings of Elispot
and/or CD57+ cells, so the patient should be consid
ered for a “monitoring” of the activity tests and
possible future symptoms. Otherwise there will be t
he risk of a relapse, a new infection or a co-
It should be noted that laboratory parameters alone
do not give a definite proof of Lyme disease,
but very important evidence and details about a pos
sible infection. The parameters are also very
important to make decisions about the duration and
success of therapy.
Adjoining is an overview
of relevant diagnostic
parameters of tick-borne
These may be extended
or limited individually
during an extensive
consultation meeting with
your attending physician
(e.g. in case of pre-
Based on the
established if such
additional laboratory tests
An overview of prices can
be found in the
attachment “Order for
laboratory tests with
declaration of consent“ for
Note: The laboratory analysis is only one part
of a comprehensive diagnosis. The
based on an extensive
a physical examination is the crucial part. The
experienced physicians of the cooperating
Medical Partnership know the broad range of
possible disease patterns, which can also
become noticeable as „
“ (note adjoining chart).
Generally, the laboratory diagnosis causes the foll
1. for Borrelia IgM- and IgG- antibodies including
-LTT and CD3-/CD 57+ T-lymphocytes
with blood sampling and special tubes
2. Basic laboratory (blood count, liver-kidney-clot
Possible necessary additional examination when susp
icion of co-
infections (Each test app. 61 – 97 €):
up to app. 945 €
Elispot- and CD3-/CD57+ are principally not covered
by health insurances!
The unit prices for different laboratory tests resu
lt from the laboratory order form in the attachment
blood samplings and special tubes will be billed se
Laboratory tests of possible co-infections
Laboratory tests of possible co-infections have an
increasing importance when adequate evidence
exists (BCA co-infections sheet). Especially Ehrlic
hia/Anaplasma, Babesia, Bartonella, Rickettsia,
Chlamydia and Mycoplasma are to be named.
Because: numerous symptoms of the co-pathogens over
lap the same symptoms of Lyme disease
patients. Without an exact knowledge of a patient’s
possible co-infections the therapist cannot
make a exact decision about the antibiotic therapy.
Not all co-infections can be treated by the
generally used antibiotics for Lyme disease. Howev
er, the testing for co-infections can induce
extensive additional laboratory costs (up to 945€).
But these are justified by the additional securit
in the diagnosis and especially by the correct anti
biotic decision, i.e. more success in antibiotic
therapies as well as generally less expensive costs
for medication during the antibiotic therapy
(vs. the “classical” antibiotic therapy of chronic
Apart from the LTT for Borrelia further cellular ac
tivity tests for Ehrlichia/Anaplasma, Chlamydia
pneunomiae and Chlamydia trachomatis have been deve
loped in the meantime (Babesia cellular
activity testing is coming soon) and can therefore
be determined with Elispot-LTT-technique. With
the help of this new Elispot
-LTT many activities of Chlamydia and Ehrlichia hav
e been detected
in the BCA! A serum examination concerning co-infec
tions is perfomed at the same time. There
are now well standardised antibody tests for Chlamy
dia, Mycoplasma, Ehrlichia, Bartonella,
Rickettsia, Babesia, Yersinia etc. The same applie
s here as well as with the Borrelia-LTT: the
antibodies alone do not have any significance towar
ds the activity of an infection – but the
-LTT is significant as it attests the high-specific
interferon release against the respective
co-pathogen in the blood.
It should be noted that in some cases it is the co-
pathogen itself which is responsible for the
symptoms and not the Borrelia infection: e.g. Chla
mydia cause diseases patterns such as Morbus
Alzheimer, Multiple Sclerosis, fibromyalgia, Chroni
c Fatigue Syndrome (CFS), myocardinal
infarcts, strokes, vasculitides, visual disturbance
The Lyme infection may well have been treated succe
ssfully with antibiotics, but the co-infection
might not have been destroyed by it. Therefore, an
exact anamnestic documentation of the
symptoms during the therapy is necessary before, du
ring and after a Borrelia infection.
Further information regarding the importance of co-
infections can be found in the separate
information for patients: „Increasing importance of
co-infections for Lyme disease patients” (article
by Armin Schwarzbach, M.D., PhD).
In case of a suspected tick-borne disease physician
s have the opportunity to execute specific
laboratory analyses in cooperation with the BCA. T
he blood kits contain special anamnestic
questionnaires by the BCA and a questionnaire to ch
eck for co-infections. The patient should fill
out these questionnaires and send them to the BCA t
ogether with the blood kits.
We are delighted to provide interested physicians w
ith further information about possibilities in
terms of collaboration and co-operation to provide
a successful treatment for patients suffering
from tick-borne diseases. Especially for the diagn
osis and therapy of chronically ill patients, who
are not living in the Augsburg area we can coordina
te the procedure of the treatment with the
attending physician at home if wanted. However, in
case of complicated symptoms and severe
illness patterns we recommend the physicians of the
Medical Partnership which cooperates with
the BCA for the first diagnosis and therapy plan pr
eparation. We also recommend the Lyme
Disease “Intensive Treatment and Rehabilitation” Pr
ogram of the BCA, which is a special Compact
Treatment clinic for chronically ill patients here
The BCA regularly offers seminars and workshops, as
well as exchange and sharing of
experiences to interested physicians and cooperatio
Below you will find as
further information regarding the laboratory tests
cellular level (Borrelia Elispot
-LTT, CD3-/CD57+ cells and the Elispots for Chlamyd
Ehrlichia) as well as the order form for laboratory
tests with declaration of consent for the BCA.
Further information regarding the laboratory tests
on cellular levels: Borrelia Elispot
CD3-/CD57+ cells, Chlamydia Elispot
-LTT and Ehrlichia Elispot
Order form for laboratory tests with declaration o
Determination of actual activity in the blood regar
ding Borrelia burgdorferi
A Borrelia infection simultaneously leads to an act
ivation of T-lymphocytes lateral to the humoral
immune response. The T-cellular immune answer vanis
hes as soon as the Lyme disease infection
is not active anymore.
A control of success of a Lyme infection therapy is
not possible via Borrelia antibodies, as the
“titer” or the antibodies can still be found in the
blood for years after a cured infection. Addition
in the first stage of Lyme disease (e.g. “bull ́s ey
e rash” or “summer flu” after a tick bite) antibodi
can only be detected and measured after several wee
ks or not at all.
The diagnostic gaps are suggested to be Borrelia ac
cording to the Elispot, which measures the
actual activity regarding Borrelia burgdorferi with
chronic and also acute Lyme disease infections.
The Elispot is so sensitive, that it can even detec
t a single Borrelia reactive T-cell in the blood. T
Elispot is 20- to 200-fold more sensitive than an E
LISA-test on Borrelia and is able to find 1
reactive cell under 100.000 lymphocytes.
The Elispot of Borrelia is very important for contr
olling a therapy of a chronic or acute Lyme
infection. In general the Elispot is negative appr
oximately 6 to 8 weeks after the end of a
Advantages of the Borrelia Elispot-LTT
(- as performed in the BCA -) in contrast to tradi
lymphocyte transformation tests:
The result is obtainable within 2 days (LTT: 1 – 2
The use of cell stabilising CPDA tubes means a sta
bility of 3 days for the measured cells
(LTT: Heparin blood only 24 hours) !
This offers a significant improvement of the stabil
ity of the examined cells and a fast decision-
making possibility for Lyme disease therapists to e
xtend the duration of a therapy or start a new
2 x 8.5 ml CPDA-tubes (kept at room temperature, d
Time required for analysis:
Billing according to GOÄ:
Number 3694 (3 different antigen approaches) factor
49.84 €) + number 4003 (Lymphocytes isolation) Fact
or 1.5 (34.97 €)
- total sum: 184.49 €
- Diagnosis of a chronic Lyme disease
- Diagnosis of a acute Lyme disease
- Decision-making regarding the length of therapy
- Control of therapy after a Lyme disease therapy
Borrelia CD 57+ cells
Determination of chronic activity in the blood rega
rding Borrelia burgdorferi
A chronic progression (stage III) of a Lyme infecti
on leads to a weakening of the immune system.
This is reflected by the decrease of the CD3-/CD57+
NK-cells in case of chronic Lyme disease.
The CD3-/CD57+ cells are a subpopulation of the Nat
A decrease of the CD57+ cells indicates (an untreat
ed) chronic or not sufficiently treated chronic
Lyme disease and does not appear in cases of a acut
e Lyme infection (e.g. “bull’s eye rash“ or
“summer flu“ after a tick bite).
The CD57+ cells reflect the degree of activity of a
chronic Lyme disease and decrease to a normal
level after a successful Lyme disease therapy (afte
r the end of treatment).
In contrast there is no decrease of CD57+ cells wit
h clinical similar diseases, such as Multiple
Sclerosis (MS), Systemic Lupus Erythematosus (SLE)
or an Amyotrophic Lateral Sclerosis (ALS).
In addition there is no significant fluctuation of
CD57+ cells throughout the day.
CD57+ cells are appropriate laboratory parameters i
n cases where chronic Lyme disease is
suspected and for therapy monitoring. These should
be measured parallel to the Borrelia Elispot,
which reflects the actual T-cellular activity.
Advantages of the CD57+ cells determination of Borr
1. The result is available within 2 days !
2. The use of cell-stabilising Heparin-tubes assure
s a stability of the measured NK-cells for 2
In combination with the Borrelia Elispot this resul
ts in a significant improvement of the stability of
the examined cells and a fast decision-making possi
bility for Lyme disease therapists to extend
the duration of a therapy or start a new treatment
1 x 10 ml Heparin-tube + 1 x EDTA-blood/blood coun
at room temperature, do not
Time required for analysis:
Ihr kompetenter Laborpartner
Determination of actual activity in the blood regar
Ehrlichia or Anaplasma are intracellular pathogens,
which can be found obligatory within the white
blood cells. Ehrlichia are transmitted to the human
by ticks contaminated with Ehrlichia
(approximately 6% of all ticks are contaminated wit
h the pathogen).
The symptoms of an Ehrlichia infection begin with f
lu like problems and express themselves with
strong headache, which are very often located “behi
nd the eyes”. Furthermore muscle aches and
numerous neurologic symptoms may be caused by Ehrli
chia. Rarely skin rashes on various parts
of the skin can be found, also on the palm of the h
and and soles of the feet.
An immune suppression is often the risk factor for
an Ehrlichia infection amongst others
depending on age, but also on chronic infections li
ke Lyme disease.
In the case that Ehrlichiosis (illness pattern of a
n infection with Ehrlichia) existing parallel to a
Lyme infection it is called co-infection. Accordin
g to the latest scientific literature additional co
infections with Ehrlichia should be taken into acco
unt in ca. 30% of all chronic Lyme disease
An infection with Ehrlichia leads to an activation
of the T-lymphocytes parallel to the humoral
immune answer (Ehrlichia-IgM- and Ehrlichia-IgG-ant
ibodies). The T-cellular immune answer
vanishes as soon as the Ehrlichia infection shows n
The Elispot on Ehrlichia/Anaplasma measures the act
ual activity of this disease.
The Ehrlichia-Elispot helps with the diagnosis of t
his infection and also is a means of a successful
control during the course of the therapy.
Advantages of the Ehrlichia Elispot
The result is available within 2 days !
The use of cell stabilising CPDA tubes means a sta
bility of up to 3 days for the measured
The right choice of a specific antibiotic therapy
2 x 8.5 ml CPDA-tubes (kept at room temperature, d
Time required for analysis:
Billing according to GOÄ:
Number 3694, factor 1.5 (49.84 €) + number 4003 (ly
isolation), factor 1.5 (34.97 €), total sum 84.81 €
- Diagnosis of an infection with Ehrlichia
- Decision-making regarding the length of therapy
- Control of therapy success after an Ehrlichia-spe
Attachment: Order form for laboratory tests with de
claration of consent
This order is to be filled out by your physician a
nd has to be signed and dated by you in case of
an order to the infecto
laboratory. The costs of the respective laboratory
order result from the
following individual prices (GOÄ), plus possible si
de costs for blood sampling, blood tubes,
shipment.BCA klinikan testejä: http://www.b-c-a.de/index.php?id=97\\\\\\\%27&L=1
----------------------------------------------------------------------------------------------------------------------------Warthin-Starry -värjäys on yksi menetelmä jolla borreliabakteereita kyetään etsimään kudosnäytteistä. http://www.ncbi.nlm.nih.gov/entrez/quer ... &DB=pubmed
Pol Merkuriusz Lek. 2006 Jun;20(120):731-4.
Clinico-pathological collations in borreliosis
[Article in Polish]
* Wagner T,
* Legatowicz-Koprowska M,
* Prochorec-Sobieszek M.
Zaklad Anatomii Patologicznej, Instytutu Reumatologii w Warszawie. firstname.lastname@example.org
Borreliosis is an infectious disease caused by spirochetal microorganisms Borrelia burgdorferi transmitted by ticks. Due to versatile clinical symptomatology and many pathogenetic aspects not explained yet, diagnosis of this disease is often very difficult. Borreliosis may affect various human organs and systems such as movement system, nervous system (central and peripheral), skin, heart and vessels. One of the diagnostic methods for detection of microbial organisms in tissues is the histochemical staining by Warthin-Starry. Very important problem connected with this illness is the relationship between borrelial antigens and autoimmunity.
PMID: 17007281 [PubMed - in process]