First isolation of Borrelia sp. (Borrelia afzelii) from cerebrospinal fluid in a patient with neuroborreliosis in Croatia

Abstract

Opisan je bolesnik s lajmskom boreliozom koja se manifestirala eritemom migrans na licu mjesec dana nakon uboda krpelja te poslije jednog mjeseca razvitkom faciopareze po perifernom tipu uz limfocitni meningitis (127 stanica/μL i 78% limfocita). Klinička slika neuroborelioze bila je razmjerno blaga. Bolesnik je liječen parenteralno ceftriaksonom kroz tri tjedna, uz dobar učinak antibiotske terapije te gotovo potpuno povlačenje klijenuti ličnog živca do kraja hospitalizacije. Bolesnik je imao reaktivna protutijela IgG na B. burgdorferi u serumu. Utvrđen je pozitivan indeks intratekalne sinteze protutijela IgM i IgG za borelije (specifičan test IDEIA Lyme Neuroborreliosis Oxoid, Velika Britanija). Imunosne pretrage na krpeljni meningoencefalitis, varicela-zoster virus i herpes simplex bile su negativne u serumu i u likvoru. U Mikrobiološkom institutu Medicinskog fakulteta u Ljubljani obra|en je likvor specifičnim uzgojnim metodama te je kultivirana B. burgdorferi sensu lato. Poslije kultivacije uzročnika izvršena je identifikacija borelijske vrste restrikcijom cjelokupnog borelijskog genoma enzimom MluI čime je utvrđen uzročnik bolesti: Borrelia afzelii. Premda se u Hrvatskoj neuroborelioza dijagnosticira i liječi preko dvadesetak godina, borelija dosada nije bila izolirana iz cerebrospinalnog likvora u naših pacijenata.We describe a patient with lyme borreliosis that manifested with facial erythema migrans one month after tick bite and consequently with peripheral facial paresis with lymphocyte meningitis (127 cells/μL and 78 % lymphocytes). Clinical picture of neuroborreliosis was relatively mild. The patient received ceftriaxone parenterally for three weeks, with beneficial effect of antibiotic therapy and almost complete resolution of facial paresis until discharge from hospital. Reactive IgG antibodies to B. burgdorferi were found in the patient’s serum. A positive index of intrathecal synthesis of IgM and IgG antibodies to borreliae was determined (specific test IDEIA Lyme Neuroborreliosis Oxoid, Great Britain). Immunological examination for detection of tick-borne meningoencephalitis, varicella-zoster virus and herpes simplex proved negative in both serum and cerebrospinal fluid (CSF). B. burgdorferi sensu lato was isolated from CSF using specific culturing techniques at the Institute of Microbiology and Immunology of the Medical Faculty, University of Ljubljana. After cultivation of the pathogen, identification of Borrelia species was performed by using restriction enzyme MluI thus determining the causative agent of neuroborreliosis: Borrelia afzelii. Although neuroborreliosis has been diagnosed and treated for over twenty years in Croatia, so far Borrelia has never been isolated from cerebrospinal fluid in our patients