Anaplasma phagocytophilum has been first isolated from the blood of two Czech patients simultaneously with a cultivation of Borrelia burgdorferi sensu lato from their erythema migrans lesions. Cultivation of different Borrelia spp. from 12 erythema migrans biopsies, from 2 blood, one liquor and one placenta sample in BSK-H medium was successful. Adapted conventional methods targeting 16S rRNA and OspA genes for real-time polymerase chain reaction (PCR) and partial sequencing of these genes together with microscopical examinations of the blood smears provided a direct detection of the B. afzelii, B. burgdorferi, B. garinii, B. valaisiana and B. bissettii in the skin, B. garinii in the blood, placenta and liquor in 24 (36.3 %) patients, and A. phagocytophilum in 10 (15 %) patients with erythema migrans. Positive indirect IgM immunofluorescence against Anaplasma sp. was obtained in 7 cases, specific IgG antibodies were detected in 12 patients. Three women suffering from erythema migrans in the first trimester had positive PCR for Anaplasma and/or for Borrelia in the blood and two of them, later, in the placenta. Interpretation of laboratory data can bring important contribution to establishing the role of Anaplasma sp. in erythema migrans and forming the principle of precaution with laboratory diagnosis during pregnancy which always should be reflected in the resistance of Anaplasma sp. toward penicillins.

• MeSH
• Anaplasma phagocytophilum immunology   isolation & purification   MeSH
• Antigens, Surface genetics   MeSH
• Bacterial Vaccines genetics   MeSH
• Borrelia burgdorferi genetics   immunology   isolation & purification   MeSH
• Diagnosis, Differential MeSH
• Adult MeSH
• Ehrlichiosis blood   diagnosis   microbiology   MeSH
• Erythema Chronicum Migrans blood   diagnosis   microbiology   MeSH
• HL-60 Cells MeSH
• Pregnancy Complications, Infectious diagnosis   microbiology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lipoproteins genetics   MeSH
• Lyme Disease blood   diagnosis   microbiology   MeSH
• Adolescent MeSH
• Placenta microbiology   MeSH
• Bacterial Outer Membrane Proteins genetics   MeSH
• Antibodies, Bacterial blood   MeSH
• Aged MeSH
• Pregnancy MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Aged MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, Surface MeSH
• Bacterial Vaccines MeSH
• Lipoproteins MeSH
• OspA protein MeSH Browser
• Bacterial Outer Membrane Proteins MeSH
• Antibodies, Bacterial MeSH

During the period 2000-2003, patients hospitalized for suspected tick-borne encephalitis in the Czech Republic were screened for possible A. phagocytophilum co-infection. Blood samples taken at admission were tested for the presence of A. phagocytophilum DNA by nested PCR using a modified target sequence as an internal control, and sera were tested for the presence of antibodies by indirect immunofluorescence and western blotting methods using cell-culture-derived antigens. To verify the assay specificity, a set of 45 sera of Patagonian residents served as a non-tick-exposed control group, and a set of 14 B. henselae-positive sera was used to check cross-reactivity. Of 809 patients hospitalized, 80 (9.9%) showed IgG antibodies reactive to A. phagocytophilum at > or =80 (reciprocal dilution factor) and 50 (6.2%) at > or =160; two (0.2%) patients showed elevated IgM titers of 40. No full blood obtained from 162 patients tested positive in PCR when false negativity was excluded. During hospitalization, the diagnosis of tick-borne encephalitis was confirmed in 536 patients, 57 (10.6%) of whom had anti-A. phagocytophilum IgG antibodies reactive at > or =80 and 41 (7.6%) at > or =160, which did not differ significantly from the whole set (P = 0.66/0.30), the maximum IgG titer registered was 5120, and no IgM titer reached the 40 cut-off. Available paired sera from 189 tick-borne encephalitis patients showed no significant shifts, but one case of slight seroconversion (IgG shift from < 80 to 320) was detected in one of the non-tick-borne encephalitis patients. The sex of the patient showed no significance for the prevalence of A. phagocytophilum antibodies; however, the seropositive patients were older on average than those who were seronegative (43.5 +/- 15.9 vs. 37.9 +/- 18.3 years, P = 0.05). Clinical manifestation of the disease did not differ noticeably between patients with and without A. phagocytophilumreactive antibodies, except for fever duration, which was significantly longer in patients with titers > or =1280. Overall, A. phagocytophilum co-infection did not seem to be a frequent and/or significant complication of tick-borne encephalitis acquired in the Czech Republic.

• MeSH
• Anaplasma phagocytophilum * genetics   MeSH
• Child MeSH
• DNA, Bacterial analysis   MeSH
• Adult MeSH
• Ehrlichiosis complications   diagnosis   epidemiology   immunology   MeSH
• Fluorescent Antibody Technique, Indirect MeSH
• Immunoglobulin G blood   immunology   MeSH
• Encephalitis, Tick-Borne complications   diagnosis   epidemiology   immunology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Polymerase Chain Reaction MeSH
• Child, Preschool MeSH
• Antibodies, Bacterial blood   immunology   MeSH
• Aged MeSH
• Case-Control Studies MeSH
• Age Factors MeSH
• Blotting, Western MeSH
• Cross Reactions MeSH
• Check Tag
• Child MeSH
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Child, Preschool MeSH
• Aged MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Geographicals
• Czech Republic epidemiology   MeSH
• Names of Substances
• DNA, Bacterial MeSH
• Immunoglobulin G MeSH
• Antibodies, Bacterial MeSH