Determination of antibodies (Humoral Immune System, Serology)

Determination of Antibodies

A direct confirmation of pathogens causing an infectious illness would be the best (3) but is not always possible and depends on the technical conditions at hand.

Indirect test procedures that measure the patient’s immune response can be used instead. In doing so, we observe what actions the immune system is undertaking against the pathogen. This works under the condition that the immune system has indeed reacted and that the test procedures are sensitive and specific enough to measure this immune response. In these considerations, the interactions between the pathogens (not only Borrelia) and the immune system of the person concerned should be observed with (2).


The humoral and the cellular immune system

There are, if we may say roughly, two areas of the immune system to be observed. On the one hand, there is the part of the immune system that produces the antibodies (the humoral immune system) and the cellular immune system on the other hand which gives us additional and useful information.

To evaluate the activity of the humoral immune system, we measure activity, quantity and type of antibodies (serological tests, e.g. ELISA, CLIA, immunoblots). The Elispot is used to test the activity on the cellular level.


Discrepancies in test methods

It is important to mention the fundamental problem of general accepted standardisation. All laboratories doing the test on blood or liquor samples do not work with a standardised antibody system, but various and different test systems. As a consequence, the same blood and liquor samples may result in different findings (negative, borderline or positive). The deviations of the results of the same sample (that can reach a factor 8 in liquor samples, (1)) depend mainly on the sensitivity of the testing system but also on the experience of the laboratory. And these problems still exist until now like Dr. Brian Fallon showed (see the new study below, 5).

This means that an evaluation on the efficiency of such tests, and therefore their results, cannot be assessed with certainty.  Existing antibodies may not be found if the test system uses inappropriate antigens. In other words, the patient might have antigens but they are not recognized by the test procedure. Every physician should only know this.

Neither laboratories, nor physicians can be “blamed” for this. After all, it is not the responsibility of an attending physician to ensure appropriate and sufficiently sensitive test procedures. He trusts the laboratory results. The point is not to blame anyone but to inform about the real difficulty this situation shows for everybody involved.

Generally, the following should be asked: Do we look at the right “place” (sample) with the right “means” (antigens) for the right pathogens? Even according to the general medical opinion, it is important to use appropriate and sensitive test procedures but how “good” or “bad” is the sensitivity in the currently used tests?

The duration and manifestation of the illness is, of course, relevant, but must be accompanied by meaningful tests.

If there was an antibody test procedure with sufficient sensitivity, then there should be an official recommendation so that all laboratories may apply the same test procedures and provide us with more security and comparison.

Unfortunately, the laboratory results are usually the only foundation for a diagnosis. Therefore, the diagnosis of an infection, carried out by the attending clinical doctor, generally has to be confirmed based on those laboratory results. Negative results (e.g. due to insufficiently sensitive test procedures) sometimes also lead to a complete exclusion of such infection. Is that right?

It is worth asking how we can find a unified and scientific foundation for discussions, if there is no standard yet for laboratory tests and that a diagnosis for an infection depends on uncertain findings of antibodies? A deviation of factor 8 (such as found in the results of liquor samples in Prof. Reiber’s study) indeed sparks questioning about the quality of laboratory results (1). Up to now, the general opinion is that only the processes and evaluation of antibodies (and the levels) can be used for a diagnosis (and activity) of an infection. However, this has changed. Today, even the general opinion agrees with the fact that those antibody processes are not sufficiently significant for the activity of this incfection, at least not in case of borreliosis (or Lyme disease), and that new parameters shall be found. Regrettably, we only know what does not seem appropriate, but no alternatives to laboratory parameters, especially for the chronic form or “late form”, have been suggested.


Progress of antibody titer

It was believed that the progress of antibody titer alone can be decisive in the measurement of the activity of the infection and its treatment. Meanwhile, it is common knowledge that the progress of antibody titer is not sufficient to measure the activity of borrelias. New parameters need to be found. The progress of antibody titer is not appropriate to control the progress of the treatment. Everyone agrees to this nowadays.

Unfortunately, we only know what is NOT appropriate but alternatives and new laboratory parameters are not offered, especially in the case of the chronic stage of Lyme. We hope to found a new parameter to measure progress using the cellular tests.


How important is the clinical picture?

The clinician (or attending practitioner) must decide what importance those tests have in his diagnosis. The laboratory values are an auxiliary.

The clinical picture (i.e. the symptoms) plays an important role. Indeed, we are giving greater importance to the patient’s symptoms as long as there are no “better” test procedures available. Actually, laboratory evidence for other pathologies is often sufficient, e.g. for “seronegative” rheumatoidarthritis. Sometimes, no laboratory evidence is needed to diagnose pathology, such as for the diagnosis of MS, schizophrenia, burnout, fibromyalgia, depression, Alzheimer or Parkinson`s. Why is it not possible to have a “seronegative” borreliosis? Why is it not enough to have clinical symptoms? Naturally, we would also prefer to be able to prove the existence of antibodies in the patient’s blood, but it implies that there are efficient tests available.

And maybe, as mentioned in the chapter “Symptoms”, the seronegative RA and borreliosis are in fact chlamydosis and/or yersiniosis.

Plus, there is the newly as human-pathogenic discovered borrelia strain, borrelia miyamotoi, which cannot yet be detected by our current test procedures. Consequently, the affected patients might receive negative results on common antibody tests (see also General Information).  Maybe there are more undiscovered and human-pathogenic strains.

Another discussed issue: could it not be possible that the IgM-Persistance is a sign of a persistent or reactivated infection, and because of this persistence, not only a marker for the early or acute stage?

An additional question: could it be that there are also problems with the sensitivity and specificity of the antibody- test- systems for other and co-infections (and not only for borrelia) and that these infections can’t all be detected  in the patients? (please see above and literatur (2), (4) )


Note: Fortunately 1-2 test manufacturers react now in 2015 with the development of antibodies testing systems with improved sensitivity ! Apparently the need for action has been noted after all. But the most test manufactures react not. However, on the other hand, this means that the sensitivity of testing systems up to now possibly has not been as good as it previously had been reported in many reviews and reports?

At the present the antibodies tests probably cannot deliver information about the activity of the infection. But the answer to this question about the activity of the infection is very important for the doctor and the patient. Hereto celluar immunology tests could eventually help. Until now there is one standardised celluar testing system or technique, this is the EliSpot technique respectively the Interferon-Gamma test (see next chapter).




(1)   Prof. Hansotto Reiber, Instand Ringversuch für Borreliose: “Liquordiagnostik und Qualitätskontrolle, CSF und Complexity Studies”, Sao Paolo, Brasil; Instand e.V. 22.11.2013

Comment: Laboratory results of the same sample of cerebrospinal fluid (CSF) vary up to factor 8!  So far, only samples for the acute neuroborreliosis were tested.  For chronic neuroborreliosis, there is no data yet. Question: Should we absolutely exclude neuroborreliosis if the used laboratory test could not find a positive result? Or could it be that the used laboratory test system was simply unable to detect a positive result?


(2)  McManus M, Cincotta A:” Effects of Borrelia on host immune system: Possible consequences for diagnostics.” Adv Integr Med (2015),


(3) Yoon E, Vail E, Kleinman G, Lento PA, Li S, Wang G, Limberger R, Fallon JT, Lyme disease: A case report of a 17-year old male with fatal Lyme carditis, Cardiovascular Pathology (2015), doi: 10.1016/j.carpath.2015.03.003


(4) Matsuda K. “A novel therapeutic strategy for mycoplasma infectious diseases”, Personalized Medicine Universe (2015),                                                                                                                                                                                     Comment: In this study the author reported also about the problems in the antibodie-tests for mycoplasma-infections because there are different mycoplasma strains, not only mycoplasma pneumoniae.



(5) A new study: Brian A. Fallon et al.: “A comparison of Lyme Disease Serologic Test results from 4 Laboratories in patients with persistent symptoms after antibiotic treatment”,  Clinical Infectious Diseases, 2014:59 (12): 1705-1710:                                                                                                                                                                         Conclusions of this study: “Although there was surprisingly little difference among the laboratories in percentage of positive results on most assay using CDC criteria, interlaboratory varability was considerable and remains a problem in Lyme Disease testing”.


(6) Vayssier-Taussat Muriel et al: Identification of Novel Zoonotic Activity of Bartonella spp., France; Emerg Infect Dis. 2015 Mar [date cited].

Further Studies about Insensitivity of some Antibody-Tests-Systems (ELISA, Immunoblot) in the blood:


  • Wojciechowska-Koszko I, Mączyńska I, Szych Z, et al. (2011): Serodiagnosis of Borreliosis: indirect immunofluorescence assay, enzyme-linked immunosorbent assay and immunoblotting. Arch Immunol Ther Exp (Warsz) 59(1), 69-77


  • Durovska J, Bazovska S, Ondrisova M, et al. (2010): Our experience with examination of antibodies against antigens of Borrelia burgdorferi in patients with suspected lyme disease. Bratisl Lek Listy 111(3), 153-5.


  • Ang CW, Notermans DW, Hommes M, et al. (2011): Large differences between test strategies for the detection of anti-Borrelia antibodies are revealed by comparing eight ELISAs and five immunoblots. Eur J Clin Microbiol Infect Dis 30(8), 1027-32




Autoren / Studien



1993 Schmitz et al.: Eur J Clin Microbiol Infect Dis 1993; 12:419-24

66 %

100 %

1995 Engstrom J Clin Microbiol 1995;33:419-27

55 %

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1996 Ledue TB, Collins MF, Craig WY: J Clin Microbiol 34, 2343-50

44 %

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1999 Trevejo RT, Krause PJ et al.: J Infect Dis 179, 931-8

29 %

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2001 Nowakiwski et al.: Clin Infect Dis 33, 2023-27

66 %

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2003 Bacon RM, Biggerstaff BJ et al.: J Infect Dis 187, 1187-99

67 %

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2005 Coulter P, Lema C et al.: J Clin Microbiol. 43(10), 5080-84

25 %


2008 Steere AC, Mc Hugh G et al.: Clin Infect Dis 47, 188-95

18 %

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2008 Binnicker MJ, Jesperson DJ et al.: J Clin Microbiol 46, 2216-21

49 %

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2009 Klemann W, Huismans BD, Umwelt-Medizin-Gesellschaft 22(2), 132-138

60 %



Further Reading