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Križna reaktivnost bakterij Treponema denticola in Borrelia burgdorferi sensu lato
Avtor: Rok Juric, stud. stom.
Mentor: prof. dr. Ana Zlata Dragaš, dr.med
Somentor: as.dr. Eva Ružic-Sabljic, dr.med, as.dr. Katja Seme, dr.med
UVOD. Bakterije reda Spirochaetales, v katerega spadata tudi rodova Treponemae in Borreliae, so monofileticnega izvora in imajo skupne nekatere morfološke, antigenske in patogenetske znacilnosti. Zaradi antigenskih podobnosti med njimi obstaja verjetnost za križno reaktivnost, ki je lahko vzrok za napacno razlago rezultatov seroloških testov pri spirohetozah.
NAMEN. V naši raziskavi smo želeli oceniti stopnjo križne reaktivnosti med skupino bakterij Borrelia burgdorferi sensu lato, povzrociteljico lymske borelioze, in bakterijo Treponema denticola, najpogosteje izolirano ustno spiroheto v zobnem plaku pri bolnikih z zmerno in napredovano parodontalno boleznijo.
MATERIALI IN METODE. Bakterijo Treponema denticola ATCC 35404 (T. denticola) smo kultivirali na NOS gojišcu pri anaerobnih pogojih. Iz tako gojenih treponem smo pripravili antigen za indirektni imunofluorescencni test (IFT), s katerim smo ocenjevali stopnjo reaktivnosti med spiroheto T. denticola in IgM oz. IgG protitelesi proti bakteriji B. burgdorferi (serumi bolnikov s sumom na lymsko boreliozo, n=303), protitelesi proti ustnim spirohetam (serumi bolnikov s parodontalno boleznijo, n=5) in protitelesi proti T. pallidum (serumi bolnikov s sifilisom, n=5). Kontrolno skupino predstavljajo serumi bolnikov z etiološko potrjeno drisko, ki niso preboleli nobene od spirohetoz (n=50).
REZULTATI. Optimizirali smo pogoje za rutinsko kultivacijo spirohete T. denticola na NOS gojišcu v našem laboratoriju. Ugotovili smo statisticno znacilno pozitivno korelacijo med vrednostmi IgM in IgG proti B. burgdorferi oz. T. denticola v IFT testu (IgM: p=0.253, p<0.005; IgG: p=0.692, p<0.001), zaradi cesar lahko z manj kot 0.5% tveganjem trdimo, da obstaja križna reaktivnost med bakterijama B. burgdorferi sensu lato in T. denticola ATCC 35404. Za oceno križne reaktivnosti med T. denticola in drugimi bakterijami (ustne spirohete, T. pallidum) ni bilo zadostnega števila serumov.
ZAKLJUCKI. Zaradi križne reaktivnosti med spirohetama B. burgdorferi in T. denticola je potrebna kriticnost pri interpretaciji pozitivnih rezultatov seroloških preiskav lymske borelioze, še posebej, kadar klinicna slika borelioze ni znacilna in tipicna. Takrat je potrebno s parodontalnim pregledom ovreci ali potrditi možnost lažno pozitivnih rezultatov zaradi prisotnosti parodontalne bolezni
[Abstract / English version]
Cross-reactivity of bacteria Treponema denticola and Borrelia burgdorferi senu lato
Author: Rok Juric, stud. stom.
Mentor: prof. dr. Ana Zlata Dragaš, dr.med
Co-mentor: as.dr. Eva Ružic-Sabljic, dr.med, as.dr. Katja Seme, dr.med
INTRODUCTION. Genera Treponemae and Borreliae are members of Spirochaetales bacterial group, which is of monophyletic origin. Members of Spirochaetales share some of morphologic, antigenic and pathogenic features. Antigenic similarities between these bacterias increase the possibility for their cross-reactivity which could be the cause for false positive results and wrong interpretation of serological tests for spirochetal diseases.
PURPOSE. The aim of our study was to estimate the cross-reactivity between Borrelia burgdorferi sensu lato, causative bacteria of Lyme disease, and Treponema denticola, which is one of the most often isolated oral spirochaetes from dental plaque among patients with moderate to advanced periodontitis.
MATERIALS AND METHODS. T. denticola ATCC 35404 was cultivated on NOS medium anaerobically. These treponemas were used as whole cell antigens in indirect fluorescent-antibody staining (IFA) procedure. IFA was used to assess a reactivity between T. denticola and antibodies of IgM and IgG class against B. burgdorferi (serum samples of persons suspected to have Lyme disease, n=303), antibodies against oral spirochaetes (serum samples of periodontitis patients, n=5) and antibodies against T. pallidum (serum samples of patients with veneral syphilis, n=5). Control group consisted of serum samples from pateints with ethiological confirmed diarrhaea but no history of any spirochaetal diseases (n=50).
RESULTS. In our study we optimised conditions for the routine cultivation of T. denticola on NOS medium in our laboratory. We found a statisticaly significant positive correlation between values of IgM and IgG antibodies against B. burgdorferi and T. denticola (p=0.253, p<0.005 and p=0.692, p<0.001, respectively). Results of our IFA studies indicated a significant positive correlation between B. burgdorferi sensu lato and T. denticola ATCC 35404 (p<0.005). Lack of serum samples unabled us to assess the reactivity between T. denticola and other bacterias (oral spirochaetes, T. pallidum).
CONCLUSION. Since our study demonstrated significant cross-reactivity between B. burgdorferi and T. denticola, there is a need of criticism in interpretation of positive results of Lyme disease serological tests especially when clinical signs and symptoms of Lyme disease are not typical. In these cases periodontal status survey could help in assessment of possibily false positive results because of the presence of periodontal disease.