Bartonella serology for patients with intraocular inflammatory disease.

PURPOSE: To determine the role of Bartonella henselae in intraocular inflammatory disease and identify its clinical features. METHODS: We retrospectively determined the serum immunoglobulin (Ig)G and IgM antibodies against B. henselae and Bartonella quintana by enzyme immunoassays in stored sera of 138 consecutive newly referred patients with uveitis who, during the acute stage of their ocular disease, underwent a standardized screening protocol to determine the cause of uveitis. RESULTS: For the entire series, the frequency of high positive levels of IgG (above 1:900) or IgM (above 1:300) antibody against B. henselae was 6% (8/138) and 3% (4/138), respectively. Except for cross-reactions between B. henselae and B. quintana, we did not find additional evidence for cross-reactions among the various bacteria tested (Coxiella burnetii and Chlamydia pneumoniae). All patients with proven infectious uveitis (n = 21) and those with established uveitic entities (n = 37) had negative B. henselae serology. High positive IgG levels were observed in 9% of patients (5/54) with unknown cause of uveitis, in two subjects with human leukocyte antigen (HLA)-B27 positive uveitis, and in one with sarcoidosis. Five patients with uveitis of unknown origin and highly elevated IgG levels against B. henselae exhibited clinical features characterized by papillitis with surrounding retinal focal lesions or edema. CONCLUSIONS: The serologic and clinical data indicate that uveitis in seropositive cases may be caused by B. henselae. We do not recommend including testing for B. henselae in initial screening of patients with uveitis, but consider it worthwhile for those with papillitis and screening results within normal limits.

Pitfalls and fallacies of cat scratch disease serology: evaluation of Bartonella henselae-based indirect fluorescence assay and enzyme-linked immunoassay.

The diagnostic value of the detection of immunoglobulin G (IgG) and IgM by Bartonella henselae-based indirect fluorescence assay (IFA) and enzyme-linked immunoassay (EIA) for the diagnosis of cat scratch disease (CSD) was evaluated. The IFA was performed either with B. henselae that was cocultivated for a few hours with Vero cells or with noncocultivated B. henselae as the antigen. Additionally, the performance of a Bartonella PCR hybridization assay based on the 16S rRNA gene was determined and compared with those of the serologic assays. The study group consisted of 45 patients suspected of suffering from CSD by fulfilling one or more of the classical criteria. The specificities of the immunoassays were set at > or = 95% by analysis of sera from 60 healthy blood donors. It is shown that the sensitivities of the IgG assays are very low (40.9% for the IFA with noncocultivated B. henselae as antigen) and that those of the IgM assays are higher (71.4% for the EIA) for patients who fulfilled two or more criteria for CSD. The IgM EIA showed the highest sensitivity: 71.4% in patients with two or more criteria for CSD and 80.6% for patients with a positive Bartonella PCR result. The results indicate that the specificities of both IFA and EIA IgG serologies and the sensitivity of the IFA IgM serology need to be improved. The PCR hybridization assay showed a sensitivity of 86.4% for patients who fulfilled two or more criteria for CSD and 100% for seven patients who fulfilled three or more criteria. The kinetics of IgG and IgM antibody production were studied in 18 patients with CSD on the basis of a positive B. henselae IFA IgM serology. The results indicate that there is no standard course of anti-B. henselae IgG and IgM production in patients with CSD, because some patients produced high levels of both IgG and IgM, others produced only high levels of IgM, and a few patients produced only low levels of antibodies.

Comparison of two panels of monoclonal antibodies for determination of Chlamydia trachomatis serovars.

A panel of monoclonal antibodies was developed for serovar typing of clinical isolates of Chlamydia trachomatis. The panel could distinguish all 15 established serovars from one another, although the hybridomas of the panel were developed by fusions of myeloma cells and spleen cells from mice immunized with antigen derived from the urogenital serovars D to L3. The typing assay was based on a dot enzyme immunoassay, and the monoclonal antibodies that were included in the panel reacted strongly in this assay. A collection of 289 clinical isolates from The Netherlands was typed. The observed serovar frequency distribution was 51 isolates of serovar D (17.6%), 103 isolates of serovar E (35.6%), 62 isolates of serovar F (21.5%), 28 isolates of serovar G (9.9%), 14 isolates of serovar H (4.8%), 2 isolates of serovar I' (0.7%), 20 isolates of serovar J (6.9%), and 9 isolates of serovar K (3.1%). These results were confirmed by typing these isolates with a panel of monoclonal antibodies purchased from the Washington Research Foundation, Seattle. No strain variation was observed within serovar D with both panels. However, restriction fragment length polymorphism analysis of the gene encoding the major outer membrane protein showed that 32 isolates were similar to the prototype D and 17 were similar to the variant D-. The two others showed a new restriction pattern. Our panel of monoclonal antibodies contained one monoclonal antibody that divided the serovar G isolates into two groups. This differentiation was confirmed by restriction fragment length polymorphism analysis, confining this difference to a known sequence variation in variable domain IV. These data support the subdivision of serovar G into serovars G (prototype strain UW-57) and Ga (prototype strain IOL-238).

An enzyme immunoassay to detect specific antibodies to protein and lipopolysaccharide antigens of Chlamydia trachomatis.

We have developed an enzyme immunoassay to measure antibodies to the proteins and lipopolysaccharide (LPS) of Chlamydia trachomatis. Antibodies to proteins could be differentiated from antibodies to lipopolysaccharide (LPS) by treatment of the antigen with periodate or Triton X-100. Some important parameters of the oxidation by periodate were studied by comparing the response of several monoclonal antibodies. Four types of response could be observed: type I, a reduced response after mild or strong oxidation; type II, a normal response after mild oxidation, but reduced after strong oxidation; type III, not affected; type IV, an increased response after oxidation. Treatment with Triton X-100 had the same effect as mild oxidation and confirmed the response types I, III, and IV. Treatment of antigen with periodate reduced the IgG response measured in sera from patients with evidence of Chlamydia psittaci infection.