Eva Ružić-Sabljić

Publications (15)33.63 Total impact

• Article: In vitro susceptibility of European human Borrelia burgdorferi sensu stricto strains to antimicrobial agents.

  Gorana Veinović, Tjaša Cerar, Franc Strle, Stanka Lotrič-Furlan, Vera Maraspin, Jože Cimperman, Eva Ružić-Sabljić
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  ABSTRACT: Broth microdilution and macrodilution assays were used to determine minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of six antimicrobial agents (ceftriaxone, cefuroxime sodium, azithromycin, amoxicillin, doxycycline and amikacin) for nine European human isolates of Borrelia burgdorferi sensu stricto (s.s.). Strains were obtained from patients diagnosed with Lyme borreliosis in Slovenia. Modified Kelly-Pettenkofer medium with a final inoculum of 10(5)Borrelia cells/mL and incubation periods of 72h and of 3 weeks and 6 weeks were used in the determination of MICs and MBCs, respectively. Observed MICs indicated that all isolates were susceptible to all the tested antimicrobial agents with the exception of amikacin. Cefuroxime sodium (MIC(90)=0.063mg/L), azithromycin (MIC(90)=0.22mg/L) and ceftriaxone (MIC(90)=0.25mg/L) displayed the lowest MICs, followed by amoxicillin (MIC(90)=1mg/L) and doxycycline (MIC(90)=2mg/L); no strain was susceptible to amikacin (MIC(90)=256mg/L). MBCs after incubation for 3 weeks and 6 weeks were determined for amoxicillin (MBC(90)=32mg/L), doxycycline (MBC(90)=32mg/L) and amikacin (MBC(90)=1024mg/L) and were found to be high (but not defined) for azithromycin (MBC(90)>0.88mg/L), cefuroxime sodium (MBC(90)>4mg/L) and ceftriaxone (MBC(90)>4mg/L). In determination of borrelial susceptibility to antimicrobial agents, intrinsic low susceptibility or methodological factors could result in low in vitro susceptibility of individual strains. This study is the first report on the antibiotic susceptibility of a series of European human isolates of B. burgdorferi s.s.
  International journal of antimicrobial agents 01/2013; · 3.03 Impact Factor
• Article: Solitary erythema migrans in children: comparison of treatment with clarithromycin and amoxicillin.

  Tea Nizič, Eva Velikanje, Eva Ružić-Sabljić, Maja Arnež
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  ABSTRACT: To compare clinical effectiveness and safety of treatment with clarithromycin and amoxicillin in children with solitary erythema migrans (EM). Consecutive patients younger than 15 years, referred to our institution in 2004 and 2005 with previously untreated solitary erythema migrans, were included in this prospective study. Basic demographic features and clinical data were collected by means of a questionnaire. The efficiency of treatment of acute disease, development of later major and/or minor manifestations of Lyme borreliosis (LB), and side effects of treatment were surveyed by follow-up visits during the first year after the initiation of antibiotic treatment. The study included 68 female and 67 male children patients. The median age of the patients was 6.5 years. Out of 135 patients, 66 received clarithromycin and 69 amoxicillin. Before treatment no differences in demographic and clinical characteristics between the two groups were observed. The mean duration of EM after the beginning of treatment was 4 days in both groups. Associated symptoms during treatment were present for 7 days in patients treated with clarithromycin and for 10 days in patients receiving amoxicillin (p = 0.188). Minor manifestations of LB were identified in 11 (22.0 %) of 50 patients receiving clarithromycin, and in 16 (29.6 %) of 54 patients receiving amoxicillin who remained in the study during the entire observation period. Major manifestations of LB were not identified in any patient treated with clarithromycin, while there were 2 (3.7 %) patients with major manifestations of LB, who were receiving amoxicillin. Side effects of treatment were identified in 24.2 % patients receiving clarithromycin and 28.1 % patients treated with amoxicillin (p = 0.761). Presence of the Jarisch-Herxheimer's reaction at the beginning of treatment was comparable in both groups (10.6 % and 10.3 %;p = 0.823). Clarithromycin and amoxicillin are equally effective and safe in treatment of children with solitary EM and have comparable side effects.
  Wiener klinische Wochenschrift 07/2012; 124(13-14):427-33. · 0.81 Impact Factor
• Article: Protein profile determination of Borrelia afzelii and Borrelia garinii isolated from skin and cerebrospinal fluid

  Urška Glinšek, Tina Udovič, Tjaša Cerar, Franc Strle, Eva Ružić-Sabljić
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  ABSTRACT: In Europe the initial skin manifestation of Lyme borreliosis (erythema migrans) is predominantly caused by Borrelia afzelii while nervous system involvement is usually associated with Borrelia garinii. The aim of our study was to compare protein profiles of B. afzelii and B. garinii isolated from skin of patients with erythema migrans and from cerebrospinal fluid (CSF) of patients with Lyme neuroborreliosis. We analyzed 187 Borrelia strains, 74 B. afzelii and 113 B. garinii, for the presence of flagellin, and outer surface proteins A, B and C. Their protein profiles were obtained by SDS–PAGE and analyzed by computer program Gel Doc. Differences in the presence of proteins were found comparing isolates according to species (B. afzelii and B. garinii) and to their origin (skin and CSF isolates); the heterogeneity regarding the presence or absence of borrelial proteins was established within particular species. Protein profiles of the analyzed strains showed differences in the number, amount and molecular mass of analyzed proteins. Distinctions in the synthesis of outer surface proteins may play a role in the dispersal of borreliae within and between animal reservoir and vector ticks, as well as in pathogenesis of Lyme borreliosis in humans.
  World Journal of Microbiology and Biotechnology 04/2012; 25(7):1287-1296. · 1.53 Impact Factor
• Article: Pregnant women with erythema migrans and isolation of borreliae from blood: course and outcome after treatment with ceftriaxone.

  Vera Maraspin, Eva Ružić-Sabljić, Dušica Pleterski-Rigler, Franc Strle
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  ABSTRACT: The course and outcome in 7 pregnant European women with erythema migrans and borreliae isolated from blood, treated with intravenous ceftriaxone for 14 days, were uneventful, and the outcome of their pregnancies was good. Spirochetemia was not associated with constitutional symptoms.
  Diagnostic microbiology and infectious disease 09/2011; 71(4):446-8. · 2.45 Impact Factor
• Article: Borrelia burgdorferi sensu lato bacteremia in Slovenian children with solitary and multiple erythema migrans.

  Maja Arnež, Eva Ružić-Sabljić
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  ABSTRACT: To establish the frequency and characteristics of Borrelia burgdorferi sensu lato bacteremia in Slovenian children with solitary and multiple erythema migrans, 1164 patients were included in this prospective study. Bacteremia was established in 11.4% of all patients, 15.8% of patients with multiple erythema migrans, and in 7.6% of patients with solitary erythema migrans. Bacteremia can be detected in children, with untreated erythema migrans, up to 39 days after the onset of skin rash.
  The Pediatric Infectious Disease Journal 06/2011; 30(11):988-90. · 3.58 Impact Factor
• Article: Acute myositis associated with the initial phase of tick-borne encephalitis.

  Maja Arnež, Tatjana Avšič-Županc, Eva Ružić-Sabljić
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  ABSTRACT: We present a case of clinically overt acute myositis associated with the initial phase of tick-borne encephalitis in a ten-year-old boy. The diagnosis of the disease was confirmed by detection of tick-borne encephalitis virus RNA in blood prior to the development of serum specific antibodies and the second phase of the disease.
  Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 06/2011; 51(4):276-8. · 3.12 Impact Factor
• Article: Comparison of erythema migrans caused by Borrelia burgdorferi and Borrelia garinii.

  Franc Strle, Eva Ružić-Sabljić, Mateja Logar, Vera Maraspin, Stanka Lotrič-Furlan, Jože Cimperman, Katarina Ogrinc, Daša Stupica, Robert B Nadelman, John Nowakowski, Gary P Wormser
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  ABSTRACT: A comparison of patients with erythema migrans due to Borrelia garinii versus Borrelia burgdorferi has not been reported. One hundred nineteen patients from New York State with erythema migrans caused by B. burgdorferi were compared with 116 patients from Slovenia with erythema migrans due to B. garinii infection. Patients with B. garinii infection were older, more often reported a tick bite, and developed larger lesions (median largest diameter: 18 and 14  cm, respectively; p = 0.01) that more often had central clearing (61.2% compared with 35.3%; p < 0.0001). Patients infected with B. burgdorferi, however, more often had systemic symptoms (68.9% vs. 37.1%; p < 0.0001), including fatigue (p < 0.0001), arthralgia (p = 0.0003), myalgia (p < 0.0001), headache (p = 0.0008), fever and/or chills (p < 0.0001), and stiff neck (p < 0.0001), and more often had abnormal physical findings (57.1% compared with 11.2%; p < 0.0001), such as fever (p = 0.0002) or regional lymphadenopathy (p < 0.0001). There was a trend for more patients with B. burgdorferi infection to have multiple erythema migrans skin lesions (13.4% compared with 5.2%; p = 0.051), and among patients with multiple erythema migrans the number of lesions was greater in B. burgdorferi-infected patients (median: 5.5 compared with 2.0; p = 0.006). The results of the present study indicate that in patients with erythema migrans the clinical features vary according to whether infection is caused by B. garinii or B. burgdorferi.
  Vector borne and zoonotic diseases (Larchmont, N.Y.) 05/2011; 11(9):1253-8. · 2.61 Impact Factor
• Article: Loss of plasmids of Borrelia burgdorferi sensu lato during prolonged in vitro cultivation.

  Urška Glinšek Biškup, Franc Strle, Eva Ružić-Sabljić
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  ABSTRACT: In the present study we analyzed stability of plasmid content in 34 Borrelia strains of three different species (13 Borrelia afzelii, 10 Borrelia garinii and 11 Borrelia burgodorferi sensu stricto) using pulse field gel electrophoresis (PFGE). During long-term in vitro cultivation consisting of 50 passages, plasmid loss was established in 46% of B. afzelii, 40% of B. garinii and 36% of B. burgdorferi sensu stricto strains. Loss of plasmids occurred as early as between the 5th and 10th passage, affected only plasmids in the range 9-41 kb but not plasmids in the range 50-68 kb and manifested with the loss of one to up to three plasmids.
  Plasmid 03/2011; 66(1):1-6. · 1.52 Impact Factor
• Article: Comparison of post-Lyme Borreliosis symptoms in erythema migrans patients with positive and negative Borrelia burgdorferi sensu lato skin culture.

  Daša Stupica, Lara Lusa, Tjaša Cerar, Eva Ružić-Sabljić, Franc Strle
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  ABSTRACT: Limited data exist on differences of erythema migrans patients with either positive or negative Borrelia burgdorferi sensu lato skin culture. We analyzed 252 adult patients with erythema migrans in whom skin biopsy specimen was cultured for the presence of B. burgdorferi sensu lato. Evaluations of epidemiological, clinical, and microbiological findings were conducted at baseline, 14 days, 2, 6, and 12 months after treatment with either doxycycline or cefuroxime axetil. One hundred fifty-one (59.9%) patients had positive skin culture (86.9% B. afzelii, 8.0% B. garinii, 5.1% B. burgdorferi sensu stricto) and 101 (40.1%) had negative skin culture. Patients in the culture-positive and culture-negative groups were comparable for the basic demographic, epidemiological, clinical, and laboratory characteristics at presentation. Statistically significantly worse selected treatment outcome parameters in the culture-positive group compared with the culture-negative group were established during follow-up. Treatment failure was documented in two patients who were culture positive and in none in the culture-negative group. Although findings for the pretreatment characteristics were comparable between the erythema migrans skin culture-positive and culture-negative patients, some parameters indicate that borrelia skin culture positivity may predict a less-favorable treatment outcome.
  Vector borne and zoonotic diseases (Larchmont, N.Y.) 11/2010; 11(7):883-9. · 2.61 Impact Factor
• Article: Evaluation of real-time PCR targeting hbb gene for Borrelia species identification.

  Jana Ferdin, Tjasa Cerar, Franc Strle, Eva Ruzić-Sabljić
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  ABSTRACT: Several molecular methods have been employed for Borrelia species identification. Newly developed technology, real-time polymerase chain reaction (RT-PCR), combines simultaneous amplification, detection and differentiation of strains in one PCR run. The aim of the study was to perform and evaluate RT-PCR for Borreliaburgdorferi sensu lato species identification. Borrelia species identification was accomplished on 374 Borrelia strains using two approaches: 1.) MluI restriction of entire borrelial chromosome (MluI-large restriction fragment patterns, LRFP), and 2.) RT-PCR targeting hbb gene and specific melting temperature (Tm) detection. The results of the two molecular methods were compared. With MluI-RFLP we were able to differentiate all Borrelia species and their subtypes within particular species. RT-PCR based on Tm determination identified unique strains within the species Borreliaafzelii (Tm 66.11 degrees C), B. burgdorferi sensu stricto (Tm 68.18 degrees C), Borreliaspielmanii (Tm 59.45 degrees C) and Borreliavalaisiana (Tm 59.62 degrees C). We were not able to distinguish the last two species that shared almost identical Tm. The large majority of Borreliagarinii strains shared Tm 51.42 degrees C, while subtype Mlg4 was characterized by Tm 56.87 degrees C. Strains of Borrelialusitaniae species also were heterogeneous; human isolate had Tm 63.47 degrees C while two tick isolates shared Tm 61.77 degrees C. Differences inside hbb gene enabled differentiation of the majority of Borrelia species, and revealed two clusters within B. garinii and B. lusitaniae species, respectively, but it was not possible to distinguish B. spielmanii form B. valaisiana. The major advantage of RT-PCR was that it was easy to perform and that the results were obtained within a few hours.
  Journal of microbiological methods 05/2010; 82(2):115-9. · 2.43 Impact Factor
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  Article: Lyme borreliosis and acute peripheral facial palsy in Slovenian children.

  Maja Arnez, Eva Ruzić-Sabljić
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  ABSTRACT: To determine how often Slovenian children with acute peripheral facial palsy are infected with Borrelia burgdorferi sensu lato, 52 patients with peripheral facial palsy were included in this prospective clinical study. According to case definitions, the diagnosis of Lyme borreliosis was established in 56% of those patients. The diagnosis was confirmed in 41%, probable in 28%, and possible in 31% of patients.
  The Pediatric Infectious Disease Journal 12/2009; 29(2):182-4. · 3.58 Impact Factor
• Article: Characterization of Borrelia burgdorferi sensu lato isolates by pulsed-field gel electrophoresis after MluI restriction of genomic DNA.

  Eva Ruzić-Sabljić, Anamarija Zore, Franc Strle
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  ABSTRACT: At least three Borrelia species (Borrelia afzelii, Borrelia garinii and Borrelia burgdorferi sensu stricto) cause disease in humans, but Borrelia spielmanii, Borrelia valaisiana, Borrelia lusitaniae and Borrelia bissettii have also been reported to be rare or potential causes of human disease in Europe. Pulsed-field gel electrophoresis after MluI restriction of the genomic DNA (MluI large restriction fragment patterns, LRFPs) represents one of several approaches that have been used to assess Borrelia genotypic characteristics. The aim of the present report was to analyze the value of MluI-LRFP for identification of B. burgdorferi sensu lato at a species level and for further species subtype delineation. Results of the present study are based on 1487 B. afzelii strains, 285 B. garinii strains, 29 B. burgdorferi sensu stricto strains, 23 B. valaisiana strains, 8 B. spielmanii strains and 3 B. lusitaniae strains. Using MluI-LRFP, we were able to delineate all Borrelia species included in the study. Each of the six examined Borrelia species displayed unique MluI-LRFPs that enabled straightforward separation of strains into particular species, and also of strains within species. The subtypes of B. afzelii (Mla2 and Mla3), B. spielmanii (Mls1 and Mls2) and B. lusitaniae (Mll1 and Mll2) uncovered in the present analysis have not been reported previously. MluI-LRFP represents a highly specific and reproducible method for Borrelia identification.
  Research in Microbiology 07/2008; 159(6):441-8. · 2.76 Impact Factor
• Article: Clinical assessment of patients with suspected Lyme borreliosis.

  Katarina Ogrinc, Eva Ružić-Sabljić, Franc Strle
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  ABSTRACT: The aim of the study was to evaluate patients with suspected Lyme borreliosis, who were referred to the Lyme borreliosis Outpatients' Clinic, Dept. of Infectious Diseases, University Medical Centre Ljubljana, Slovenia, for examination. All adult patients with suspected Lyme borreliosis (with the exception of those having erythema migrans) seen at our Outpatients' Clinic by one of the authors of this report (K.O.) between January 2003 and November 2006 were included in the study. Demographic, epidemiological, and clinical data were obtained using a structured questionnaire. Patients were classified as having probable, possible, or past Lyme borreliosis, history of a tick bite or a toxic reaction after tick bite or insect bite/sting, presence of borrelial antibodies in serum without any symptoms, or another disease not attributed to borrelial infection at all. Out of 339 patients included in the study 13 (3.8%) were considered as having probable, 59 (17.4%) as possible, and 53 (15.6%) as past Lyme borreliosis; four (1.2%) patients were assessed because of a past tick bite, one (0.3%) for a past toxic reaction after an insect sting, while three (0.9%) were tested positive for Borrelia burgdorferi sensu lato but reported no complaints. Two-hundred and six (60.8%) patients were assumed to have a distinct disease not due to borrelial infection. Comparison of patients considered as having probable/possible Lyme borreliosis with those whose complaints were interpreted as being not due to borrelial infection revealed relatively few clear distinctions. In conclusion, over 60% of patients referred to and assessed for suspected Lyme borreliosis were probably having symptoms due to another disease. Only 72 out of 339 (21.2%) patients were interpreted to have probable or possible Lyme borreliosis and were recognized as candidates for antibiotic treatment.
  International journal of medical microbiology: IJMM 06/2008; · 2.80 Impact Factor
• Article: Infection of small mammals with Borrelia burgdorferi sensu lato in Slovenia as determined by polymerase chain reaction (PCR).

  Anamarija Zore,, Miroslav Petrovec,, Katarina Prosenc,, Tomi Trilar,, Eva Ružić-Sabljić,, Tatjana Avšič-Županc
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  ABSTRACT: Thirty-four small mammals collected in the vicinity of Ljubljana were tested for the presence of Borrelia burgdorferi sensu lato by polymerase chain reaction (PCR) of urinary bladder tissues, using universal flagellin primers and species specific rRNA primers. Seventeen small mammals (50%) were found to be positive, and 7 small mammals were infected with two species of B. burgdorferi sensu lato simultaneously. The most commonly found species was B. afzelii (n = 14), followed by B. burgdorferi sensu stricto (n = 7) and B. garinii (n = 3), as determined by species-specific primers. We conclude that PCR is a rapid and reliable method to detect infection with B. burgdorferi sensu lato in small mammals.
  Wiener klinische Wochenschrift 01/2000; 111(22-23):997-9. · 0.81 Impact Factor
• Article: Comparison of different Borrelia burgdorferi sensu lato strains for detection of immune response in patients with erythema migrans

  Lilijana Merljak Skočir, Eva Ružić-Sabljić, Vera Maraspin-Čarman, Stanka Lotrič-Furlan, Mateja Logar, Franc Strle
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  ABSTRACT: The aim of the present study was to establish which combination of serological method and Borrelia strain used as an antigen would provide the most appropriate demonstration of borrelial infection in patients with eythema migrans residing in Slovenia. Four different strains were chosen as antigens: two strains of B. afzelii and two strains of B. garinii which differed in their expression of the outer proteins OspA, OspB and OspC. Each individual strain was used as antigen in immunofluorescence test (IFT), enzyme-linked immunosorbent assay (EIA) with whole borrelial cells, and EIA with ultrasonicated borrelial cells. With these 12 different tests, 100 samples were examined for the presence of specific IgM and IgG antibodies: 50 sera of blood donors and 50 sera of patients with erythema migrans. The latter were further subdivided into skin culture-positive and -negative subgroups. A commercial Western blot (WB) test was performed for 26 sera of the control group and 25 sera of patients with erythema migrans. The four different methods had distinct specificity and sensitivity. The most specific approaches were IFT (100% for IgM and 90–92% for IgG) and the WB test (100% for IgM and 73% for IgG), followed by EIA with whole borrelial cells (80–98% for IgM and 76–84% for IgG) and EIA with ultrasonicated borrelial cells (76–94% for IgM and 72–80% for IgG). The sensitivity levels of all these tests were low. The most sensitive were EIA tests with whole borrelial cells (28–36% for IgM and 32–42% for IgG) followed by EIA with ultrasonicated borrelial cells (22–32% for IgM and 24–36% for IgG), the WB test (16% for IgM and 32% for IgG) and IFT (0–2% for IgM and 14–20% for IgG). The following methods gave significant differences between patients and negative controls in detecting IgM antibodies: EIA with whole borrelial cells with both B. afzelii antigens and with antigen B. garinii that expressed OspA and OspC, EIA with ultrasonicated borrelial cells with antigen B. afzelii that expressed OspA, OspB and OspC. In detecting IgG antibodies, significant differences were observed between EIA with whole borrelial cells and with antigen B. afzelii that expressed OspA and OspB. Borreliae were isolated from the skin of 34/50 (68%) patients with erythema migrans: two strains failed to grow, while 26/32 (81%) strains were identified as B. afzelii, 5/32 (16%) as B. garinii and 1/32 (3%) as B. burgdorferi sensu stricto. No statistically significant differences in serologic test results between culture-positive and -negative patients with erythema migrans were found.
  International Journal of Medical Microbiology.