Bela P. Bozsik, M.D.

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.
Memberships:
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 (Wikipatents, USPTO ). Read more about dr. Bozsik's method below and here.
????: Founder, Med, Sec. Head of the Hungarian Lyme Borreliosis Foundation, Budapest (http://lymenet.hu). 

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!

"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! 

"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: 

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