Serodiagnosis of borreliosis: indirect immunofluorescence assay, enzyme-linked immunosorbent assay and immunoblotting.

Lyme disease is an infectious, multi-system, tick-borne disease caused by genospecies of Borrelia burgdorferi bacteria sensu lato, characterized by remarkable heterogeneity. In this situation choosing an optimal antigen array for diagnostic tests seems problematic. The serological tests for borrelia routinely done in laboratories often produce ambiguous results, which makes a proper diagnosis rather complicated and thus delays the implementation of an appropriate treatment regimen. Thirty-seven outpatients and eight inpatients with suspected borreliosis diagnosis hospitalized at the Clinics of the Pomeranian Medical University (Szczecin, Poland), participated in the study. In order to detect the antibodies against Borrelia sensu lato three kinds of serological tests were used: indirect immunofluorescence assay (IIFA), enzyme-linked immunosorbent assay (ELISA), and immunoblot. The IIFA and immunoblot tests conducted on 45 patients (100%) produced positive results for both the IgM and IgG antibody types. In the case of ELISA, positive or borderline results were observed in only 24 patients (53.3%). The immunoblot test for IgM most frequently detected antibodies against the outer surface protein C (OspC) antigen (p25), and, in the case of IgG, against the recombinant variable surface antigen (VlsE). The IIFA screening test used for diagnosing Lyme borreliosis produced the highest percentage of positive results, which were then confirmed by immunoblot, but not by ELISA. Therefore using only ELISA as a screening test or for diagnosing Lyme borreliosis seems debatable.

Evaluation of the level of antibodies against Chlamydophila (Chlamydia) pneumoniae in post-surgery heart ischaemia patients and their clinical conditions - a six-year study.

INTRODUCTION: Inflammatory conditions modulated by Chlamydophila (Chlamydia) pneumoniae are considered to play an important role in the onset of atherosclerosis. In this paper we present the results of progressive observation of C. pneumoniae antibody titres in patients who underwent coronary artery bypass graft (CABG). MATERIAL AND METHODS: The objective of our research was a prospective observation of antibody titres in IgA and IgG class antibodies against C. pneumoniae using indirect immunofluorescence in a group of 155 post-surgery CABG patients suffering from heart ischaemia. The microbiological test results were compared with patients' present coronary complaints evaluated on the CCS scale during a six-year period. RESULTS: Six years after CABG, 128 patients (82.6%) are still alive. During the study a positive serological conversion of antibody titres was observed in 36 patients in the IgA class antibodies, and in 26 patients in the IgG class. The group of patients with no antibodies against C. pneumoniae decreased from 23.2 to 3.4%, while the group of patients with antibodies in both IgG and IgA classes increased from 52.3 to 83.9%. The average CCS degree decreased from 3.18 before CABG to 1.65 in the present study. CONCLUSIONS: These results show no connection between the serological symptoms of chronic C. pneumoniae infection and coronary complaints evaluated on the CCS scale during a six-year study on post-CABG patients suffering from heart ischaemia. The surgical treatment of heart ischaemia brought about long-term improvement in the coronary condition of the observed group of patients.

Antibiotic susceptibility and molecular characterisation of Proteus mirabilis isolates in hospitals from the West Pomeranian area of Poland.

Proteus mirabilis isolates (n = 177), collected between 1996 and 2000 in four hospitals in the West Pomeranian area of Poland, were characterized by antibiotype and pulsed-field gel electrophoresis (PFGE). The selected isolates were collected from different wards (intensive care unit, surgery, internal medicine, and urology). The strains were cultured from various specimen types, mostly from urine, wound samples, bronchial exudates and sputa. The identification was done by biochemical test ID 32E ATB (bioMerieux). Analysis of PFGE patterns was based on comparison of the banding patterns obtained by PFGE of chromosomal DNA digested with SfiI enzyme. Among all P. mirabilis isolates tested three major genotypes A (A1-A7), B (B1-B4), C (C1-C5) and 71 unique patterns were identified. The same genotypes were obtained from different patients, treated in different wards and hospitals during a 5-year period. The strains which belonged to the genotypes A and B were multiresistant and most of them produced ESBL; genotype C was more sensitive to antibiotics.

Proinflammatory cytokine (IL-1beta, IL-6, IL-12, IL-18 and TNF-alpha) levels in sera of patients with subacute cutaneous lupus erythematosus (SCLE).

Subacute cutaneous lupus erythematosus (SCLE) is a subset of lupus erythematosus that identifies patients with clinically recognized erythematous, nonscarring lesions, photosensitivity and serologic abnormalities. Anti-Ro (SS-A) antibodies are considered to be a typical immunopathologic marker of SCLE. Autoimmune diseases have been also characterized by the disturbances in the cytokine network. The aim of this study was to compare the concentrations of proinflammatory cytokines (IL-1beta, IL-6, IL-12, IL-18 and TNF-alpha) in serum of ANA-positive (antibody against nuclear antigen) and ANA-negative patients with SCLE. Sera samples were collected from 15 patients with SCLE (9 ANA-positive and 6 ANA-negative ones). The preliminary identification of autoantibodies as well as their titers was determined on HEp-2 cells using IIF method. Western blotting (EUROIMMUN) was applied to verify the results of IIF. Proinflammatory cytokine concentrations in the patients' sera samples were determined by enzyme-linked immunosorbent assay (ELISA) (Bender MedSystems). The levels of IL-12 were higher in ANA-positive patients than in ANA-negative subgroups [median (interquartile range), 330 pg/ml (128-708 pg/ml) versus 39.4 pg/ml (31.25-80 pg/ml)]. Similar differences were observed in the level of IL-18 [median (interquartile range), 508.4 pg/ml (180-1222 pg/ml) versus 100.5 pg/ml (78.1-154 pg/ml)]. The differences in TNF-alpha levels between the groups of ANA-positive and ANA-negative patients were at the verge of statistical significance, p<0.05. The sera levels of IL-1beta and IL-6 were low and of no significant difference concerning the ANA-positive and ANA-negative subgroups. Since serum levels of IL-12 and IL-18 were higher in ANA-positive patients than in ANA-negative patients, these cytokines might play an important role in the inflammatory process in SCLE.