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© Borgis - New Medicine 3/2010, s. 84-87
*Małgorzata Tokarska-Rodak1, Hanna Fota-Markowska2, Maria Kozioł-Montewka1, Filip Śmiechowicz2, Roma Modrzewska2
The detection of specific antibodies against B. burgdorferi s.s., B. afzelii , B. garinii, and B. spielmani antigens in patients with Lyme disease in eastern Poland
1Department of Medical Microbiology, Medical University, Poland
Head of Department of Medical Microbiology: prof. dr hab. Maria Kozioł-Montewka
2Department of Infectious Diseases, Medical University, Lublin, Poland
Head of Department of Infectious Diseases: prof. dr hab. n. med. Roma Modrzewska
Summary
Aim. The purpose of the investigation was to determine the presence of antibodies for B. burgdorferi s.s., B. afzelii , B. garinii , and B. spielmani in persons with Lyme disease from the Lubelski region (eastern Poland) in the aspect of existing disease symptoms.
Materials and methods. The study was conducted in a group of 45 patients with the second stage of Lyme disease hospitalized in 2007-2008. The presence of anti- B. burgdorferi s.l. antibodies IgM and IgG was determined in the patients' serum (Western blot test, Mikrogen).
Results. The antibodies (IgM/IgG) for four gene-species, B. burgdorferi s.s., B. afzelii , B. garinii , and B. spielmani, were present in 39 patients from 45 tested. The presence of IgM anti-OspC antibodies for 1 gene-species, B. spielmani, was found only in 1 person.
Symptoms involving bone/joint structure in the tested group of patients were present among 41 persons, and symptoms of muscle pain among 21. Patients also complained of headaches and attention deficit disorder (6 persons). Erythema migrans appeared in 2 persons.
Conclusions. The results of the study suggest the possibility of co-infections with several gene-species of B. burgdorferi s.l. evidenced by the presence of IgM antibodies to OspC of B. burgdorferi s.s., B. afzelii , B. garinii , and B. spielmani.The detection of antibodies directed against specific B. spielmani antigens suggests that this microorganism may be responsible for triggering Lyme disease both as a single etiologic agent and in co-infection with other Borrelia gene-species. A relation between the types of symptoms present among patients with Lyme disease and gene-species responsible for causing diseases was not found.
Introduction
The complex of Borrelia burgdorferi sensu lato encompasses at least 12 species: B. burgdorferi sensu stricto, B. garinii , B. afzelii , B. spielmani , B. valaisiana , B. lusitaniae , B. japonica , B. andersonii , B. tanukii , B. turdae , B. bissettii and B. sinica. The main pathogenic genomic species responsible for human Lyme disease in Central Europe and Eastern Europe are: Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii. However, the DNA of B. valaisiana, B. lusitaniae , and B. spielmani detected in samples of human origin or the spirochetes were isolated from patients with symptoms of Lyme disease (1, 2). The genetic variability of Borrelia species has a considerable impact on the pathogenesis and clinical picture (3, 4, 5, 6). The heterogeneous clinical manifestation observed in Lyme disease inclined researchers to analyze the relation of infection by particular gene-species of B. burgdorferi s.l. with clinical symptoms that appeared. Borrelia afzelii seems to be associated with acrodermatitis chronica atrophicans (ACA) and with borrelia lymphocytoma, B. garinii with neuroborreliosis and B. burgdorferi s.s with arthritis (2, 3). Patients with EM reveal infection with all three gene-species: B. burgdorferi s.s., Borrelia afzelii and Borrelia garinii. Examinations of cerebrospinal fluid in patients with neurological symptoms of Lyme disease have revealed the possibility of a relation of neuroborreliosis with the presence of each of those three gene-species (7). DNA of B. valaisiana was detected in the cerebrospinal fluid of a patient with chronic neuroborreliosis in Greece and in a patient with erythema migrans. B. lusitaniae was isolated from a patient with suspected Lyme disease in Portugal (3). A direct association of skin changes of erythema migrans (EM) type with an infection of B. spielmani was revealed in some European countries (Netherlands, Germany, Hungary, Slovenia). Thus, the association of these gene-species with Lyme disease has been proven (8). The number of substantiated infections of B. spielmani is still insufficient in order to present the differences in Lyme disease manifestations caused by B. spielmani in comparison with Lyme disease caused by other gene-species. To date, an investigation which could explicitly affirm the relation of Lyme infection morbidity with infection by B. spielmani within Poland has not been performed yet.
Aim of the study
The aim of the study was to determine the presence of antibodies for four gene-species of Borrelia burgdorferi s.l. - B. burgdorferi s.s., B. afzelii , B. garinii , and B. spielmani - in persons with Lyme disease from the Lubelski region (eastern Poland) in the aspect of existing disease symptoms.
Materials and methods
The study was conducted in a group of 45 patients: 23 men (age 20-66 yrs) and 22 women (age 20-60 yrs) with the second stage of Lyme disease hospitalized in the Department of Infectious Diseases, Medical University of Lublin in 2007-2008.
The study group involved patients demonstrating manifestations of joint involvement including arthralgia and/or arthritis treated symptomatically and receiving etiotropic antimicrobial therapy. The diagnosis of borreliosis was established on the basis of the patient's medical history, physical examination, clinical picture, and serologic investigation (ELISA and Western blot).
Testing for antibodies IgG and IgM to Borrelia burgdorferi sensu lato was performed using the Western blot test (recomLine Borrelia IgG and IgM, Mikrogen) which included antigens for IgM and IgG: OspC ( B. burgdorferi s.s., B. afzelii , B. garinii , B. spielmani), p18 ( B. burgdorferi s.s., B. afzelii , B. garinii , B. spielmani), VlsE, p100, p58, p41, p39, OspA. The presence of anti- B. burgdorferi sensu lato antibodies IgM and IgG was determined in the patients' serum.
Results
IgM antibodies alone against specific Borrelia burgdorferi s.l. antigens were detected in 17 of 45 patients (38%). The IgM antibodies anti-OspC and anti-p18 B. burgdorferi s.l. were observed with varying frequency in this group. Table 1 shows IgM antibodies anti-OspC and anti-p18 B. burgdorferi s.s., B. afzelii , B. garinii, and B. spielmani in persons with Lyme disease.
Table 1. IgM antibodies against B. burgdorferi s.l. specific antigens in patients with Lyme borreliosis.
N=17Presence of IgM anti-B. burgdorferi s.l. antigens
anti-OspCanti-p-18anti-p41
12b, a, g, s-+
1b, a, g, sb, s+
2a, g, s-+
1b, a, s-+
1a, s-+
b – Borrelia burgdorferi sensu stricto
a – Borrelia afzelii
g – Borrelia garinii
s – Borrelia spielmani
+ presence of antibodies
IgM and IgG antibodies against specific Borrelia burgdorferi s.l. antigens were detected in 28 of 45 patients (62%). Table 2 shows IgM and IgG antibodies anti-OspC and anti-p18 for B. burgdorferi s.s., B. afzelii , B. garinii , and B. spielmani in persons with Lyme borreliosis.
Table 2. IgG antibodies against specific Borrelia burgdorferi s.l. antigens in patients with Lyme borreliosis.
N=28Presence of IgM anti-B. burgdorferi s.l. antigensPresence of IgG anti-B. burgdorferi s.l. antigensPresence of IgM/IgG
anti-OspCanti-p-18anti-OspCanti-p18against other specific antigenic B. burgdorferi s.l protein
8b, a, g, s-b, a, g, s-+
4b, a, g, s---+
1b, a, g, s-aa+
3b, a, g, s--a+
1b, a, g, s---+
1b, a, g, sb, g, sa, g1, g2+
1b, a, g, s-b, a-+
1b, a, g, ssb, a, g, s-+
1b, a, g, ss-g2+
1b, a, g, s-g, s-+
1b, a, g, s-b-+
1b, a, g, s-b, g, s-+
1a, ss-a+
1b, a, g, s-ab+
1s---+
1b, a, ssg, s-+
b – Borrelia burgdorferi sensu stricto
a – Borrelia afzelii
g – Borrelia garinii
s – Borrelia spielmani
+ presence of antibodies

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Piśmiennictwo
1. Ruderko N et al.: Molecular detection of Borrelia bissettii DNA in serum samples from patients in the Czech Republic with suspected borreliosis. FEMS Microbiol Lett 2009; 292, 274-81. 2. Sicklinger M, Winecke R, Neubert U: In vitro susceptibility testing of four antibiotics against Borrelia burgdorferi: a comparison of results for the three genospecies Borrelia afzelii, Borrelia garinii and Borrelia burgdorferi sensu stricto. J Clin Microbiol 2003; 41: 1791-1793. 3. Derdakowa M, Lencakowa D: Association of genetic variability within the Borrelia burgdorferi sensu lato with the ecology, epidemiology of Lyme borreliosis in Europe. Ann Agric Environ Med 2005; 12: 165-172. 4. Wang G, Van Dam AP, Dankert J: Phenotypic and genetic characterization of a novel Borrelia burgdorferi sensu lato isolate from a patient with Lyme borreliosis. J Clin Microbiol 1999; 37: 3025-8. 5. Richter D et al.: Relationships of a novel Lyme disease spirochete, Borrelia spielmani sp. nov., with its hosts in Central Europe Appl Environ Microbiol 2004; 70: 6414-9. 6. Kurtenbach K et al.: Fundamental processes in the evolutionary ecology of Lyme borreliosis. Nature Rev Microbiol 2006; 4: 660-669. 7. Wodecka B: Borrelia burgdorferi; in: Biologia molekularna patogenów przenoszonych przez kleszcze (Ed.: Skotarczyk B), PZWL, Warszawa, 2006. 8. Maraspin V, Ruzic-Sabljic E, Strle F: Lyme borreliosis and Borrelia spielmanii. EID 2006; 12: 1177. 9. Zajkowska J et al.: Clinical forms of neuroborreliosis – the analysis of patients diagnosed in department of infectious diseases and neuroinfection medical academy in Białystok between 2000-2005. Przegl Epidemiol 2007; 61: 59-65. 10. Dybowska D, Kozielewicz D, Abdulgater A: Prevalence of borreliosis among forestry workers in kujawsko-pomorskie voivodeship. Przegląd Epidemiol 2007; 61: 67-71. 11. Garlicki A: The modern therapy of Lyme disease. Przegl Epidemiol 2007; 61: 449-456. 12. Cisak E et al.: Prevalence of Borrelia burgdorferi genospecies in Ixodes ricinus ticks from Lublin region (eastern Poland). Ann Agric Environ Med. 2006; 13: 301-6. 13. Bratton RL, Whiteside JH, Hovan MJ: Diagnosis and treatment of Lyme disease. Mayo Clin Proc 2008; 83: 566-571. 14. Wilske B, Fingerle V, Schulze-Spechtel U: Microbiological and serological diagnosis of Lyme borreliosis. FEMS Immunol Med Microbiol 2007; 49: 13-21. 15. Marangoni A et al.: A decrease in the immunoglobulin G antibody response against the VlsE protein of Borrelia burgdorferi sensu lato correlates with the resolution of clinical signs in antibiotic-treated patients with early Lyme disease. Clin Vaccine Immunol 2006; 13: 525-529. 16. Preac-Mursic V et al.: Kill kinetics of Borrelia burgdorferi are associated with distinct clinical manifestations of Lyme borreliosis. Clin Infect Dis 1993; 17: 708-717.
otrzymano: 2010-07-28
zaakceptowano do druku: 2010-08-31

Adres do korespondencji:
*Małgorzata Tokarska-Rodak
Department of Medical Microbiology, Medical University of Lublin
Chodźki Str. 1, 20-093 Lublin, Poland
phone/fax: +48 81 7423781
e-mail: rodak.malgorzata@gmail.com

New Medicine 3/2010
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