Characterization of Bartonella clarridgeiae flagellin (FlaA) and detection of antiflagellin antibodies in patients with lymphadenopathy.

Cat scratch disease (CSD) is a frequent clinical outcome of Bartonella henselae infection in humans. Recently, two case reports indicated Bartonella clarridgeiae as an additional causative agent of CSD. Both pathogens have been isolated from domestic cats, which are considered to be their natural reservoir.

Hemin-dependent growth and hemin binding of Bartonella henselae.

Bartonella henselae causes cat-scratch disease and bacillary angiomatosis peliosis. The bacteria reside in erythrocytes of asymptomatic cats, which represent the natural reservoir for this pathogen. B.

Two different genotypes of Bartonella henselae in children with cat-scratch disease and their pet cats.

Two genotypes (I and II) of Bartonella henselae are involved in cat-scratch disease (CSD). Lymph node biopsies were taken from 3 children suffering from CSD, and blood cultures were obtained from their pet cats. Cat-scratch disease was confirmed serologically, histologically and by detection of B.

Seroprevalence of antibodies to Bartonella henselae in patients with cat scratch disease and in healthy controls: evaluation and comparison of two commercial serological tests.

Serologic testing for the presence of antibodies to Bartonella henselae is a widely accepted diagnostic procedure for laboratory confirmation of the diagnosis of cat scratch disease (CSD). In this study a commercially available indirect immunofluorescence assay (IFA) based on B. henselae-infected human larynx carcinoma cells (test A) was evaluated.

Detection and identification of two Bartonella henselae variants in domestic cats in Germany.

To determine the prevalence of bacteremia caused by Bartonella henselae in domestic cats in the region of Freiburg, Germany, we investigated culture of blood from 100 cats from 89 different households over a 12-month period. B. henselae could be isolated from 13% (13 of 100) of these cats.

Comparison of serological tests for the detection of antibodies against Chlamydia trachomatis and Chlamydia pneumoniae in rheumatological patients.

In cases of reactive arthritis, a suspected Chlamydia trachomatis infection is often detected by serological methods. However, mostly tests with genus-specific antigens are used, neglecting the fact that antibodies against Chlamydia pneumoniae are highly prevalent in the adult population. Therefore we tested sera of 129 patients with various rheumatological disorders and of 18 healthy persons in parallel with a genus-specific test (IPAZYME) and with the species-specific microimmunofluorescence test for C.

Comparison of host responses after intranasal infection of guinea-pigs with Mycoplasma genitalium or with Mycoplasma pneumoniae.

Guinea-pigs were infected intranasally with Mycoplasma genitalium or Mycoplasma pneumoniae. The lung lesions produced by the two mycoplasmas were comparable in extent and histological pattern. Sera of both animal groups taken 2 weeks after infection reacted strongly in the complement fixation test with the M.

Effects of growth phase and antibiotics on quantitative DNA/RNA hybridization.

Synthetic probes complementary to ribosomal RNA are increasingly used in the detection of bacteria. Many applications, however, require the quantitation of bacteria. We therefore tested the influence of growth phase and representative antibiotics (ampicillin, chloramphenicol and gentamycin) on the outcome of DNA/RNA filter hybridization using radiolabelled probes and a multisample digital autoradiograph for quantitative monitoring.

Immunodominant epitopes of the adhesin of Mycoplasma pneumoniae.

Overlapping octapeptides from the amino acid sequence of the adherence protein of Mycoplasma pneumoniae were synthesized and used as the antigen in an enzyme-linked immunosorbent assay with serum samples from M. pneumoniae-infected patients. Of a sequence of 338 amino acids positioned between leucine 801 and leucine 1139, only two regions with immunodominant continuous epitopes were detected.

Binding sites of attachment-inhibiting monoclonal antibodies and antibodies from patients on peptide fragments of the Mycoplasma pneumoniae adhesin.

The adherence protein (P1 protein) of Mycoplasma pneumoniae was purified by electroelution and cleaved with cyanogen bromide. The resulting peptides were separated by two-dimensional electrophoresis. Spots reacting in Western immunoblots with two attachment-inhibiting monoclonal antibodies were isolated, and the amino-terminal ends of these peptides were microsequenced.

Isolation of the adherence protein of Mycoplasma pneumoniae by fractionated solubilization and size exclusion chromatography.

The 168-kDa adherence protein of M. pneumoniae was solubilized and purified to homogeneity. Optimal yield was obtained by pretreatment of whole M.

Host reactions to Mycoplasma pneumoniae infections in guinea-pigs preimmunized systemically with the adhesin of this pathogen.

Guinea-pigs developed systemic and local humoral responses after intraperitoneal immunization with the isolated adhesin (168 kDa protein) of Mycoplasma pneumoniae cells. Hilar lymphocytes of these animals showed proliferation after in vitro stimulation with the 168 kDa protein or sonicated M. pneumoniae whole cell antigen.

Antibody response of patients against a 120 kDa surface protein of Campylobacter pylori.

Campylobacter pylori strains were isolated and serum samples were obtained from 63 patients. Immunoblots of 52 patients sera using their own isolates as antigen showed a 120 kDa band, which was missing in the other 11 isolates and the respective sera. This band was not detected in other Campylobacter species.

Immunological reaction of guinea-pigs following intranasal Mycoplasma pneumoniae infection and immunization with the 168 kDa adherence protein.

Humoral responses to Mycoplasma pneumoniae proteins, especially the 168 kDa protein, were demonstrated by Western blotting in sera and bronchial washings of all groups of infected or immunized guinea-pigs. However, infection was not prevented by these local and systemic antibodies. Hilar lymphocytes of infected and immunized guinea-pigs were stimulated in vitro by sonicated M.

Antibodies in the sera of Mycoplasma pneumoniae-infected patients against proteins of Mycoplasma genitalium and other mycoplasmas of man.

Frequency and pattern of anti-Mycoplasma genitalium antibodies in sera of 50 patients with Mycoplasma pneumoniae-infection were examined by the Western immunoblot method. The sera reacted with several proteins of M. genitalium.

Amino acid sequence and antigenicity of the amino-terminus of the 168 kDa adherence protein of Mycoplasma pneumoniae.

The amino-terminal end of the 168 kDa adherence protein from the membrane of Mycoplasma pneumoniae was sequenced up to 12 amino acids. A synthetic peptide containing nine amino acids of this sequence was used to study the antigenicity of the amino-terminus of the 168 kDa protein and the involvement of the homologous sequence of the protein in the adherence process. The synthetic peptide when coupled to ovalbumin was immunogenic in rabbits.

Use of adherence protein of Mycoplasma pneumoniae as antigen for enzyme-linked immunosorbent assay (ELISA).

Antibodies against the adherence protein of Mycoplasma pneumoniae are regularly found in patients with M. pneumoniae infection. Therefore, this 168-kilodalton (kDa) protein was used as an antigen in a dot-ELISA for serological diagnosis of M.

A 168-kilodalton protein of Mycoplasma pneumoniae used as antigen in a dot enzyme-linked immunosorbent assay.

The attachment protein of Mycoplasma pneumoniae (molecular weight 168 kd) was used as antigen in a special enzyme-linked immunosorbent assay (dot ELISA) and compared with a sonicate of the whole organism. In control sera the intensity of the 168-kd band on immunoblots correlated well with the ELISA IgG values derived from isolated protein. The diagnostic significance of the 168-kd antigen was tested on paired sera from 33 patients with Mycoplasma pneumoniae infection (24 children, 9 adults).

Reaction pattern of human anti-Mycoplasma pneumoniae antibodies in enzyme-linked immunosorbent assays and immunoblotting.

In serological diagnosis of Mycoplasma pneumoniae disease the frequently used complement fixation test is based on a cross-reacting glycolipid. Recently enzyme-linked immunoassays have been developed to overcome this lack of specificity. To study the involvement of the various proteins and the influence of age on the level of antiprotein antibodies present, we investigated by enzyme-linked immunoassays (immunoglobulin M [IgM] and IgG) and immunoblotting the sera of healthy persons of different age groups as well as sera of patients (including paired sera) with M.

Mycoplasma pneumoniae induces cytotoxic activity in guinea pig bronchoalveolar cells.

Precultured guinea pig alveolar macrophages (AM) and freshly harvested alveolar cells (FHAC) activated by interaction with Mycoplasma pneumoniae were cytotoxic for xenogeneic 75selenomethionine-labeled tumor target cells. Phagocytosis of whole opsonized or nonopsonized M. pneumoniae cells was more effective in eliciting cytotoxicity than uptake of sonicated microorganisms.

Adherence inhibition assay: a specific serological test for detection of antibodies to Mycoplasma pneumoniae.

Antibodies directed against the adherence-mediating protein of Mycoplasma pneumoniae were measured by an adherence inhibition assay. Pretreatment with antibody-containing sera reduced the attachment of sheep erythrocytes to Mycoplasma pneumoniae layers grown in flat-bottom microtiter plates. The degree of attachment of erythrocytes was estimated by lysis with distilled water and measurement of absorbance in a microtiter reader.