Bela P. Bozsik, M.D.

Medical Secretary of the Hungarian Lyme Borreliosis Foundation (LBF website at; BP retired as working physician in 1999, but he continues his research in Lyme borreliosis and related infections in private clinic.
Email: bpbozsik at
Office & Postal address: Dr. Bela Bozsik, Lyme Borreliosis Foundation, Tetenyi St. 98, building B Budapest, XI Hungary H-1119

Curriculum Vitae:

Born in Budapest, Hungary, Sept. 9, 1942, as son of Pal Bozsik and Maria Klinga; married to Marta Schleer, Dec. 4, 1968; children: Bela, Andras Pal, Attila Peter.
Member of N.Y. Acad. Sci.
Member of Society for Christian Physicians, Roman Catholic.

1960-61: Worker Lenin Metall. Works, Miskolc, Hungary.
1966: Graduated as MD from Semmelweis Med. U., Budapest.
1966-74: Pathologist St. Istvan Mcpl. Hosp., Budapest.
1974-99: Researcher. Nat. Inst. Hygiene, Budapest. A new method for evaluating the immunohemolytic activity of Complement with modified method was developed and new hypothesis for the structure of Membrane Attack Complex (C5b-C9) was constructed during 1974-1984. Based on the studies of the Complement system a new micro-method for complement fixation method was elaborated.

Positions / special work and achievements regarding the subject of "Lyme borreliose" and related:
1980-99: Head of the Serology laboratory at Johan Bela Nat. Inst. Hygiene; during that periode the laboratory did over 100000 serology tests for Borrelia burgdorferi, using different methods.
1999: Head of Lyme borreliosis center, St Rokus Hosp., Budapest, a position from which he had to retire in 1999.

1985: Proposing the since then worldwide accepted technical term for Lyme Disease and Related Disorders that is "Lyme Borreliosis"at the 2nd World Conference.
????:  Proposing a diagnostic and therapeutic scheme for Lyme borreliosis seronegatives, and new hypothesis for pathogenesis of Lyme borreliosis respectively. 
2002-2003/03: Researcher at Lyme borreliosis Clinic in Budapest
2002:  Patent 6689577. Special reagent (Dualdur) and procedure for the detection of pathogens, especially spirochetes from body fluids by microscopy.

Selected excerpts from the patent description: 

"The laboratory diagnosis of borrelioses with different clinical presentations is based primarily on the detection of spirochetes from blood samples. This is easily accomplished in recurrent fever because normally, there is a large number of B.recurrentis present. Besides, other morphological properties of this pathogen (shown in the table enclosed) and the fact that this pathogen is easy to stain also make its detection easier. There are mild cases, however, when the symptoms suggest the diagnosis of recurrent fever but the cell count is too low for the conventional methods to detect the causative agent. To solve this problem, the technique of microhematocrit concentration (double centrifugation of blood samples) has been used since 1972. Microscopy is superior in that the test result is not affected by the changing antigenicity of Borreliae. [Goldsmid, J M. Mohamed: The use of Microhematrocit Technic for the Recovery of Borrelia duttonii from the Blood, Am. J. Clin. Pathol. 58:165-169 (1972)].
As mentioned above, morphological examination has long been included in the laboratory diagnosis of spirochetoses. If dark-field microscopy is employed, concentrated fluid samples do not need to be stained, which means that the long and thinspirochetes are not washed off the slides, which, in turn, increases sensitivity.

During the first stage of this infection, antibody production is slower than usual. Antibodies do not appear until weeks after the infection and are only rarely present throughout the whole course of the disease because titers keep changing and--after some time--they may become normal without any intervention. This makes it difficult to define the threshold titer. There is no clinically applicable threshold that could make a clear-cut distinction between those who are infected and those who are not. Besides, generation cycles of the causative agent cause a fluctuation of the early--IgM type--antibody titers. As far as we know, this is the only disease in which the causative agent blocks the production of the more specific and more effective IgG type antibodies, which normally follows the production of IgM. There are even cases of Lyme borreliosis in which only the early (IgM type) antibodies are present years after the infection.
What has been said so far affects all antibody assays. That is to say, comparative studies can only compare the sensitivity of the techniques in question.
Thus, it would be a big mistake to base the laboratory diagnosis of Lyme borreliosis on the traditional evaluation of a single test. Test results are sometimes considered non-specific in this case.
The chances of non-specific reactions are known to be higher in spirochetoses but they can be avoided with traditional pre-test absorption, which removes the non-specific antibodies that could give a false reaction. If the test result is negative, physicians may doubt the validity of the patients' complaints and abandon the possibility of Lyme borreliosis even though antibody production may be inadequate or blocked, the technique employed may not be able to detect all antibodies or the threshold value may not be set correctly.
In a study made at a university in Vienna, Austria in 1985, Professor Stanek and his colleagues found that in artificially infected laboratory animals bacteremia could be detected using dark-field microscopy as well as conventional microscopy after Giemsa staining: the Borrelia burgdorferi sensu lato injected subcutaneously appeared in the circulation and remained detectable continuously. The number of bacteria detected was changing in seven to eight day cycles. They realized that the number of pathogens is changing and that in certain periods of the generation cycle spirochetes are more difficult to detect. [(Stanek, G.; Burger, I; Hirschl, A.; Wewalka, G.; Radda, A: Borrelia transfer by ticks during their life cycle Studies on laboratory animals. Zbl. Bakt. Mikrobiol. Hyg. A., 263: 29-33: 1986)]
In our experience, it is still possible to detect the pathogenic bacteria if there are less than 10 bacteria in a milliliter of centrifuged native blood samples. In comparison, the threshold for the detection of Lyme borreliosis with PCR, which is currently considered the most sensitive but can only be done in specially equipped laboratories, is between 40 and 100 germs per ml; besides, as many as possible primers specific to different sub-strains should be available.
It should be noted that further morphological, immunocytological and immunoserological examination of the centrifuged sample treated with the reagent according to the invention is also possible. Furthermore, it can also be utilized for PCR and cultivation. In the latter cases, filtering is recommended before concentrating the sample."

2004: The diagnostic method (DualDur®) was Patented in USA (WikipatentsUSPTO ). Read more about dr. Bozsik's method below and here.
????: Founder, Med, Sec. Head of the Hungarian Lyme Borreliosis Foundation, Budapest ( 

Authorship of Bozsik BP on subject "Lyme borreliosis":     PubMed search for "Bozsik+BP"
1985: Borrelia burgdorferi stained with acidic acridine orange. Ann. Immunol. Hung. 1985;25:335-344
- this paper is not ref. on PubMed, but was ref. as #20 by
Woods GL, WalkerR DH. Detection of Infection or Infectious Agents by Use of Cytologic and Histologic Stains. Clin Microbiol reviews, July 1996, p. 382–404. PMID: 8809467 PDF

[Excerpt on staining of Borrelia spirochaetes: "Acridine orange, a fluorochrome dye that at low pH allows differentiation of bacteria, which show orange fluorescence, from background material and mammalian cells, which fluoresce green to yellow, is particularly useful for interpretation of smears prepared from thick or purulent material (58, 99). It also is more sensitive than the Gram stain for detection of organisms in CSF and other body fluids and in smears prepared from blood culture broth (103, 104, 126), and in at least one case it allowed detection of Borrelia burgdorferi in sediment of CSF (20*)" ... "Wright and Giemsa stains, used routinely in the hematology laboratory, demonstrate relapsing fever borrelia spirochetes in stained smears of peripheral blood."

If published in English? - as title seem to indicate - would it be possible for you to send me a photocopy, or if published in Hungarian, make abstract / comment in English?

1987: Bozsik BP, Lakos A, Budai J, Telegdy L, Ambrozy G. Occurrence of Lyme borreliosis in Hungary. Zentralbl Bakteriol Mikrobiol Hyg [A]. 1987 Feb;263(3):466-7. PMID: 3591100  No abstract available.
1990: Kristof V, Bozsik BP, Szirtes M, Simonyi J. Lyme borreliosis and Raynaud's syndrome. Lancet. 1990 Apr 21;335(8695):975-6. PMID: 1970048  No abstract available.
1995: Rules of Ticks [Hungarian].
1997: Advice on Tick Diseases [Hungarian].
2000: Bozsik BP. [Management of Lyme disease][Hungarian]. Orv Hetil. 2000 Jan 9;141(2):106-11.  PMID: 10686785  No abstract available.

Would it be possible for you to make abstract / summary in English?

2002: Bozsik BP. [Comment on Borrelia burgdorferi Group infections][Hungarian]. Orv Hetil. 2002 May 26;143(21):1223-4. PMID: 12073543 No abstract available.

Would it be possible for you to make a abstract / summary in English?

2004: Prevalence of Lyme borreliosis. Lancet. 2004 Mar 13;363(9412):901. PMID: 15031053.  No abstract available, but full text is available from The Lancet website after login with free registration!
Letter, excerpt: "Treatment with antibiotics does not always result in eradication of the organism, therefore without follow-up and repeated treatment at recurrence, Lyme borreliosis chronica can develop. Lyme borreliosis is often undetectable by serological techniques. In our practice, the passive haemag glutination method (Diagast, France) failed to detect more than 60% of cases, compared with the newer ELISA (Enzygnost, Behring, Germany). The primary and secondary errors of this passive haemagglutination method were calculated as 1·9% and 6·3%, respectively, from 50000 investigations. The significant difference between these diagnostic techniques highlights the need to assess other factors, especially clinical symptoms, in the evaluation of results and formulation of the definitive diagnosis.
The occurrence of Lyme borreliosis can be estimated from the reported incidence of tick-borne encephalitis (TBE) and the bacterial (1:10) and viral (1:1000) infectivity rate of ticks ( The estimated incidence of TBE in Hungary (population 10 million) is 200–400 cases per year, and the infectivity rate of ticks is 100 times higher for Borrelia burgdorferi sensu lato than for the TBE virus. Thus there could be more than 20000 new cases of Lyme borreliosis per year in Hungary. Given the subclinical nature of the disease, the problems with diagnosis, misunderstanding about criteria and diagnosis, and the mean age of patients being 60 years, the number of patients affected at any one time could be as much as 1million—ie, 10% of the population."

2005 Sheffield, UK, 2005 Lyme conference presentations
Read more about dr. Boszik's practical and theoretical experience gained from his many years of working with Borrelia serology and other test methods, as presented by himself in his PPT-lecture on DIAGNOSIS  (PPT) and about his experience with successful treatment of Lyme Borreliosis seronegativa (PPT)

Conclusive statements by dr. Boszik:  Based on 120000 samples / 150000 determinations since 1989: Serology test after chosing the CUT-OFF that gives smallest failures in both regards for Lyme borreliosis, will FAIL IN AT LEAST 7% of all test-results - thereof 5,5% false positives and 1,5% false negatives - but in practice serology tests do unfortunately not live up to the best theoretically possible reliability, rather fails much more often.
"The missed proportion of Lyme borreliosis seronegativa was growing with NEWER methods [PHA 60%, EIA-IgG 30%, EIA-IgM 0% (EIA Enzygnost)"
"Although 1.5% first order and 5.5% second order error are excellent results for any laboratory [but those people misdiagnosed by false positive or negative serology test results probably think it is BAD RESULT?] it is astonishing that the developments of science give possibility to determine 60% or more cases as people, who are in bad need of treatment.
This result is neither over-diagnosis, nor under-estimation especially not false/untrue determinations; they really belong to both the development of methodology and the epidemic character of Lyme borreliosis."
IT IS NOT POSSIBLE - and probably never will become possible - to safely detect NOR outrule all cases of Lyme borreliosis with one single serology test method!

Bozsik 2005 Borrelia tests comparison

"What is on C6-ELISA?  -  C6 ELISA  &  Wb  were identical in 97% at CDC comparing 180 samples .  There was not any reactivity in Master’s disease, in which B. burgd. sensu lato were isolated with substrain determination. Masters,E–Philipp,M: C6 Lyme peptide ELISA serosurvey of Missouri patients. Abstracts&Presentations at the IX Intl.Conf. on Lyme borreliosis & Abstract at The Clinical side of Lyme Disease, N.Y. Augustus 18-22., 2002."

"Closing remarks:
1. Definitive/firm diagnosis for Lyme borreliosis could be set up with laboratory methods [Dark Field microscopy + other].
2. indication of the treatment is Set up by clinician. Treatment should be followed/repeated in respect to different sensitivity of different substrains
3. by control determinations; investigations for 3-5 years."

From the Bozsik "Diagnosis" lecture, Sheffield 2005:

  1. Spirochetes could be demonstrated in BLOOD by dark-field microscopy during all ACTIVE stages of pathogenesis!
  2. 107 of 143 (75%) of the cases with live (moving) spirochetes found in their blood by dark-field microscopy,
    were confirmed by real-time PCR to belong to Borrelia burgdorferi sensu lato:
    66 (61.7%) B. burgdorferi sensu stricto
    20 (18.7%) B. garinii
     6 (  5.6%) B. afzelii and
    15 (14.0%) other Borreliae strains than the usual 3 EUROPEAN strains
    Many of these cases were also confirmed by monoclonal antibody stain for Borrelia burgdorferi with anti-ospA and anti-flagellin kindly donated by
    prof. Barbour USA.
  3. One third (1/3) of these patients with proven LATE Lyme borreliosis were SERONEGATIVE! 

Excerpt from the US patent description of DualDur®

"As the title suggests, a new reagent has been developed that can slow down the aging (membrane hardening) of human erythrocytes, leukocytes, platelets and squamous epithelial cells in the samples. It has been found that the reagent according to the invention stops the amoeboid movement of leukocytes and the fragmentation of platelets. Thus, myeloid figures are not formed. The membrane of accidentally formed myeloid figures is also hardened. Consequently, they do not even exhibit Brownian movement; they simply float along. The movement and the cell division of Borrelia burgdorferi sensu lato remained unaffected by the invented reagent. This is how shedding could be observed, which had only been noted in cell cultures." ....  "The technique according to the invention makes it possible to study the current state of pathogenesis and determine the activity of Lyme borreliosis in a given patient. A fast and reliable diagnosis can be made even when serological tests are negative. The reagent and the procedure according to the invention provides reliable data to aid treatment (which is still controversial), monitor treatment effects and predict relapses before the development of humoral immune response or after it has been blocked and all this is independent of autoimmune responses. "
.... Illustrative picture from abovementioned lecture: 

Bozsik - DualDur double centrifugation preparation method for Dark Field microscopy

Information about DualDur® copied from Wilder Network - page three:

"The results of the [DualDur®] test are not influenced by either the genetic polymorphism or the phenotype of Borrelia burgdorferi sensu lato (or the changes in either of these during the pathogenesis of the disease), which is very important in Middle-Europe. The test is also reliable in the cases when Lyme disease is caused by new subspecies or immunological changes or when vaccination has produced an antibody response. The theoretical sensitivity of this procedure can be as high as one organism/ml. Concurrent PCR tests appear to be less sensitive even nowadays. If properly evaluated, it is the most sensitive of the direct tests and it is also very reliable. It provides a tool to investigate natural phenomena in a laboratory setting, which may help to gather new information regarding the pathogenesis of Lyme disease.
In the microscopy, immobile cellular bodies are always seen in the background serving as controls of the moving Spirochetes.
DualDur® -treated normal or washed blood or Borrelia burgdorferi sensu lato cultures can also be used as controls. Indirect immunofluorescent assays using specific monoclonal antibodies kindly donated by prof. Barbour and Barbara Johnson prove that the Spirochetes are identical to Borrelia burgdorferi sensu lato. They were proven by electron microscopy in the last years with both negative staining and immuncytologic reactions.

This reagent provides conditions similar to the body, therefore the division of Spirochetes was detected several times; sometimes they were acting in pictures. You can see it in the following:

The next story will be on the emerging combined antibiotic treatments developed in 1990 and used widely with success in Hungary prescribed by the members of the Therapeutic Workgroup of the Lyme Borreliosis Foundation in Hungary. According to our practice we can tell you that the following could not be proved in Middle Europe: “Conclusion : Patients with post-treatment chronic Lyme disease who have symptoms but show no evidence of persisting Borrelia infection do not show objective evidence of cognitive impairment. Additional antibiotic therapy was not more beneficial than administering placebo." END

Bozsik & Burgdorfer
Dr. W. Burgdorfer (left), Dr. B.P. Bozsik (in the middle) and colleague?

From personal / Internet group communications with dr. Bozsik:

Bozsik's own words on Borrelia burgdorferi Bleb's & Cysts - see more pictures of Borrelia spirochetes undergoing changes in the PDF:

Please let me tell You my opinion based on my morphological practice since 1986.
There are no cyst-forms as all ball-like morphology has always some material in them. They could be differentiate according to the following :
1. Gemma􀃉 has DNA and could be further cultivated. This possibility was never proved by Garon and Dorward – Personal communication. If yes, it is the spore, until it is a sporoid structure.
2. Macromolecular immunocomplexes (blebs) contain Antigen-Antibody-Complement beside different  extracellular material and Osp-s and they never could be used for cultivations. These forms could be developed during cultivation as leading character of B.burgdorferi as other Borrelias do. They are shedding both in the body and in the cultivation media.
3. So-called Spheroplasts or L-forms are developing during the effect of cell-wall-damaging antibiotics, and Lforms are caused by a process of herniation. They could also have DNA, but there is no hypothetical possibility of
4. Further damaged structures could be detected, which are the final product of protoplasma fragmentation inside of the Spirochetes – that is the granulation. It is hard to prove their extracellular presence as the so-called coccoid forms, except DNA hybridization.
5. Different forms of the intact B.burgdorferi were demonstrated in cultivation by Aberer and Duray and those of dividing Spirochete by me.
6. Ghost could be appeared as a final state of the reorganization process of B.burgdorferi after lytic damage􀃉

Do not forget the Dictionaries:
Gemma \Gem"ma\, n.; pl. Gemm[ae]. [L., a bud.]
1. (Bot.) A leaf bud, as distinguished from a flower bud.
2. (Biol.) A bud spore; one of the small spores or buds in the reproduction of certain Protozoa, which separate one at a time from the parent cell.

1. Gemma - small asexual reproductive structure in e.g. liverworts and mosses that detaches from the parent and develops into a new individual, reproductive structure - the parts of a plant involved in its reproduction)

Detailed you can find in my lectures hold at the Anniversary Congress of Lyme Borreliosis Foundation in Budapest in 2000 about the DualDur® reagent and method and the Hypothesis based on the results got with this method. The complete summary could be downloaded at (letoltheto anyagok) and further information from now
All of my investigation was made in native human blood samples with Dark-field microscopy.
More from the conference website: Information to patients about Lyme borreliosis (10 pages, English version), Questionnaire (1 page, English version, Word DOC), Bozsik (2000) PPT on Therapy and Bozsik (2000) PPT on Diagnosis.  
PS: I have never prescribed as I am a retired pathologist, but proposed to use and our colleagues are using tinidazol with our combined treatment schedule with better effect than without it to be demolish Gemma-s. I wish to know your proposal for patients with sensitivity to “azols”. Alinia? Plaquenil? Have you practice with it?  Crossreaction with other antibiotics?  Thanks You in advance, Bela

Mail to MMI-list 11. Sept. 2007: 

Dear Colleagues and Members of the list!

Binding antibody to the protein-band of 41 means appearance of LB either in the past or in the days. Other positive binding help us to evaluate a firm positive reaction. Perhaps the formerly reagents could not avoid the cross-reaction, so there were period of time with the need of clinical differentiations. Sometimes a one-shot abx treatment causes decapitation of the immune-response and develops LB seronegativa with a weak or no reaction even to p41.  Seropositivity otherwise means only that LB exists.

The indication of the treatment belongs to the clinical symptoms at this moment. – The only absolute indication of the treatment is the PCR positivity of the biting tick. [??? why do you NOT mention PCR positive on PATIENT SAMPLE?].
The substrain(s) determination gives a good data for the personal antibiotic treatment-schedule in the Carpathian Basin and Europe, too. There are further subgroups of Borrelia burgdorferi sensu stricto with different pathogenicity, which could also differ in their sensitivity to antibiotics. 
Think of the extreme genetic plasticity and adoptability of Borrelia burgdorferi sensu lato we should prescribe combined antibiotics not allow the adoptation of the causative agent.

There is a possibility to investigate Bbsl in blood using DualDur® reagent and method to help with further data the clinicians. This reagent and method was developed in 1986.
You can see the demo video at the lower end of the page for with some words on my video:

This method gives a possibility for further analysis of Borrelia burgdorferi sensu lato in similar condition to the body.  Summarizing: there are two main and different chapters in Lyme borreliosis.

1.      Diagnosis of LB according to laboratory investigations

2.      Indication of treatment according to the activity of clinical symptoms

Of course there are several not classified parts both of diagnosis and indication of treatment, especially in the process of the development of a personal schedule for treatment.

3.      The differential diagnosis of other TBD should be introduced into these processes…

Best wishes

This article was proof read by and further comments made by Dr. Bozsik (October 2007), but some of the excerpts has been added since.