The sequences of groESL operon of Anaplasma phagocytophilum among human patients in Slovenia.

Anaplasma phagocytophilum is an emerging tick-borne pathogen. Great genetic diversity of A. phagocytophilum has been described in animals and ticks. The present study is focused on the genetic variability of the groESL operon of A. phagocytophilum in human patients in Slovenia. During 1996­2008, there were 66 serologically confirmed patients with human granulocytic anaplasmosis. Of these, 46 were tested with a screening PCR for a small part of the 16S rRNA gene of A. phagocytophilum and 28 (60.9%) were positive. Positive samples were additionally tested with a PCR targeting the groESL operon and a larger fragment of the 16S rRNA gene. All amplicons were further sequenced and analyzed. The homology search and the alignment of the groESL sequences showed only one genetic variant. Sequence analysis of the 16S rRNA gene revealed 100% identity among amplicons. Slovenia is a small country with diverse climate, vegetation, and animal representatives. In previous studies in deer, dogs, and ticks, great diversity of the groESL operon was found. In contrast, in wild boar and in human patients from this study, only one genetic variant was detected. The results suggest that only one genetic variant might be pathogenic for humans or is competent enough to replicate in humans. To support this theory, other genetic markers and further studies need to be performed.

Clinical appearance of Staphylococcus aureus spondylodiscitis and molecular characterization of the isolates.

Infectious spondylodiscitis is diagnosed with increasing frequency. It is most commonly caused by Staphylococcus aureus. The aim of the present study was to determine clinical differences between S. aureus and non-S. aureus spondylodiscitis and, by using spa typing, to determine whether certain clones of S. aureus predominate. During the y 2002-2006, 70 patients (45 male, 25 female) were diagnosed with spondylodiscitis and treated at the Department of Infectious Diseases, University Medical Centre Ljubljana. The age range was 13-95 y (mean age 63.4 y). Cultures were positive in 59 (85%) patients; S. aureus was isolated from blood and/or vertebral disc in 42 (60%) and other bacteria were isolated in 17 (25%). In patients with S. aureus spondylodiscitis, the infection was more likely to be haematogenous. These patients were more likely at risk for epidural abscess. Thirty S. aureus isolates from blood were typed with the spa typing molecular method. All of them were methicillin-susceptible. We identified 20 spa types. Seven spa types (t005, t015, t026, t091, t116, t449 and t474) were identified more than once, while 13 were sporadic. Among the sporadic spa types, 6 were newly identified in this study and introduced as new types into the spa server database. A heterogeneity of S. aureus isolates from spondylodiscitis was detected in this study. Thus no specific clone was identified that might be more likely to cause infection of the spine.