Neuroborrelios (NB) kan förekomma när Borrelia garinii, som är en del av artkomplexet Borrelia burgdorferi sensu lato (s.l), infekterar nervsystemet. När immuncellerna utsätts för borreliaantigen bildas antikroppar i plasma och intratekalt. NB ger även upphov till mononukleär pleocytos i cerebrospinalvätska (CSV). Neurologiska symptom, pleocytos och antikroppsindex (AI), som indikerar intratekal syntes av borreliaantikroppar, vägleder nuvarande NB-diagnostik på Klinisk Mikrobiologi i Kalmar. Diagnostiken försvåras vid tidig NB när AI är negativ och pleocytos positiv. För att komplettera tidig NB-diagnostik kan kemokinet C-X-C motif ligand 13 (CXCL13) mätas i CSV. I nuläget utförs CXCL13 enzymkopplad immunadsorberande analys (ELISA) på ett externt laboratorium. Mätning av CXCL13 kan även utföras med en snabb semi-kvantitativ ReaScan+ CXCL13 metod som bygger på lateral flödesimmunanalys (LFIA). Syftet var att verifiera ReaScan+ CXCL13, kontrollera linjäritet, CXCL13 stabilitet och bestämma diagnostisk specificitet och sensitivitet. CSV-proverna indelades i fyra grupper baserat på pleocytos och AI samt tre kategorier utifrån kriterier till NB-diagnos (n=31). Metodjämförelsen genomfördes mellan ReaScan+ LFIA och CXCL13 ELISA (n=25). Linjäritetsanalys utfördes på prov med hög CXCL13-koncentration i CSV och CXCL13-stabiliteten kontrollerades på två patientprover vid frys- och kylförvaring. Beräkning av sensitivitet och specificitet baserades på definitiv (n=20) och ej NB (n=5). Metodjämförelsen visade en fullständig överensstämmelse och Spearman´s korrelationskoefficienten var 0,949. Determinationskoefficienten för linjäritetsanalysen var 0,963. Diagnostisk sensitivitet var 80% och specificitet 100%. Hög prestanda, acceptabla stabilitets- och linjäritetsresultat samt ReaScan+ potentiella förmågan att särskilja mellan definitiv och ej NB, indikerar att metoden kan införas på Klinisk Mikrobiologi i Kalmar och komplettera nuvarande diagnostik av tidig NB. / Lyme neuroborreliosis (LNB) occurs when Borrelia garinii, part of the Borrelia burgdorferi sensu lato (s.l) species complex, infects the nervous system. Upon exposure, immune cells produce antibodies that can be measured in serum or cerebrospinal fluid (CSF). LNB also cause mononuclear pleocytosis in CSF. Neurological symptoms, pleocytosis and antibody index (AI), which indicate intrathecal synthesis of Borrelia antibodies, guide current diagnosis of LNB at the Clinical Microbiology in Kalmar. The diagnosis is complex in early LNB, especially when AI is negative and pleocytosis is positive. Expression of chemokine C-X-C motif ligand 13 (CXCL13) in CSF can be used as an additional marker to early LNB. Analysis is currently performed at an external laboratory using enzyme-linked immunosorbent assay (ELISA). The aim of this project was to evaluate the performance of the rapid semi-quantitative lateral flow immunoassay (LFIA) ReaScan+ CXCL13 test using 31 patient samples divided into four groups based on pleocytosis and AI. Linearity assay was performed on samples with high CXCL13 concentrations and stability was tested on two CSF samples after freezing and refrigeration. Estimation of sensitivity and specificity was based on definite (n=20) and non-LNB (n=5). Method comparison showed a complete agreement and Spearman's correlation coefficient was 0,949. The R2 for linearity was 0,963. Sensitivity was 80% and specificity 100%. Strong agreement, acceptable stability and linearity results as well as ReaScan+ potential ability to distinguish between definite and non-NB, indicate that the method can be introduced at Clinical Microbiology in Kalmar and used as a supplement in early LNB diagnostics.

During a period of 3 years, from March to July 2014, 2015 and 2016, ticks were collected from dogs and cats in shelter facilities for abandon animals in Česke Budejovice, South Bohemia. In total, 343 ticks were found on 106 pets: 67 domestic dogs and 39 cats. All collected ticks, that were identified as Ixodes ricinus and Ixodes hexagonus, were tested for the presence of spirochetes from Borrelia burgdorferi sensu lato (s.l.) complex, Rickettsia spp., Anaplasma spp, and Babesia spp using conventional PCR and nested PCR. Identification of pathogens was done by following sequencing of amplicons. Out of all tested ticks, 49,56% were proved to be infected at least with one pathogen. Co-infection of at least two different pathogens was determined in 18 ticks (5,2%). The aim of the present study was to estimate the role of accompanying animals (cats and dogs) in the circulation of ticks and tick-borne pathogens, to determine the frequency of pathogenic infections in dog and cat associated ticks, to evaluate the current risk of infection for dogs and cats, with respect torisk forhumans living in the area of České Budějovice.