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Borrelia burgdorferi, the agent of Lyme borreliosis, is genetically more heterogeneous than previously thought. In Europe five genospecies have been described from the original B. burgdorferi sensu lato (sl): B. burgdorferi sensu stricto (ss), B. garinii, B. afzelii, B. lusitaniae, and B. valaisiana. In the United States, B. burgdorferi ss as well as B. bissettii in California and B. andersonii on the East Coast were differentiated. In Asia, B. japonica has been identified along, with B. garinii, B. afzelii, and B. valaisiana. In order to evaluate sensitivity and specificity of four species-specific monoclonal antibodies, we analyzed 210 B. burgdorferi sl isolates belonging to eight genospecies by immunoblot and confirmed genospecies by restriction fragment length polymorphism (RFLP) of rrf (5S)-rrl (23S) intergenic spacer amplicon. Monoclonal antibody H3TS had 100% sensitivity for 55 B. burgdorferi ss isolates but showed reactivity with all four isolates belonging to B. bissetii. Monoclonal antibody I 17.3 showed 100% specificity and sensitivity for 45 B. afzelii isolates. Monoclonal antibody D6 was 100% specific for B. garinii but missed 1 of 64 isolates (98.5% sensitivity). Monoclonal antibody A116k was 100% specific for B. valaisiana but was unreactive with 4 of 24 isolates (83.5% sensitivity). Genetic analysis correlated well with results of reactivity and confirmed efficacy of the phenotypic typing of these antibodies. Some isolates showed atypical RFLP. Therefore, both phenotypic and genotypic analyses are needed to characterize new Borrelia isolates.

An immunoglobulin G immunoblot was developed with antigenic extracts of Borrelia burgdorferi sensu stricto, B. garinii, B. afzelii, and B. valaisiana genospecies and was reacted with sera from patients with neuroborreliosis, acrodermatitis, and Lyme arthritis. A detailed analysis of the reactivities of the protein bands was performed, and a two-step scoring procedure was selected to determine the preferential reactivity of sera to one particular genospecies. The discriminative potential of 5 proteins (12-kDa, 16-kDa, 18-kDa, OspA, and 66-kDa proteins) was used as a rapid first-step scoring method, followed by scoring of 14 additional protein bands if necessary. The advantage of this procedure is the low percentage of serum samples with inconclusive results for one of the four species (10% for patients with neuroborreliosis, 6% for patients with acrodermatitis chronica atrophicans, and 6% for patients with Lyme arthritis). Among 31 serum samples from patients with neuroborreliosis, 16 were more reactive to B. garinii, 7 were more reactive to B. afzelii, 3 were more reactive to B. valaisiana, and 2 were more reactive to B. burgdorferi sensu stricto. Of 31 serum samples from patients with acrodermatitis, 26 showed a higher level of reactivity to B. afzelii. Of 34 serum samples from patients with Lyme arthritis, 21 were more reactive to B. burgdorferi sensu stricto, 10 were more reactive to B. afzelii, and 1 was more reactive to B. valaisiana. Our results suggest an organotropism of Borrelia species and provide some evidence of a pathogenic potential of B. valaisiana in humans.

Boutonneuse fever caused by Rickettsia conorii is transmitted mainly by the brown dog tick, Rhipicephalus sanguineus. We collected 540 ticks in Marseille, France, and tried to isolate as many strains of rickettsia as possible. Ticks were evaluated for the presence of rickettsia by the hemolymph test and by a new culture system, the centrifugation-shell vial technique. We avoided contamination in the culture system. Prior to ticks being submitted to the hemolymph test, they were disinfected. Only 5.6% (27 of 478) of the cultures were contaminated. A drop of hemolymph from each of 478 R. sanguineus ticks was cultured in two shell vials, and another drop was stained by the Gimenez method or indirect immunofluorescence. Since Gimenez staining in our hands was not satisfactory, comparison of the hemolymph test and culture is based on the results of indirect immunofluorescence. Thus, 50 of 369 (13.5%) examined ticks were hemolymph test positive, and 44 (11.9%) cultures were positive. After disinfection, another pool of 62 ticks were examined by the hemolymph test. The ticks were kept individually in a sterile environment. A few days later, the hemolymph of these ticks was collected again and cultured. The contamination rate was not significantly higher (6.4%) than in the above-described conditions. It allowed us to isolate eight more strains. Thus, we recommend screening ticks with the hemolymph test and culturing only the hemolymph test-positive ticks.

From 1982 through 1987 we diagnosed 13 chronic Q fever cases. Clinically these patients presented a culture-negative endocarditis, and all but two had high complement-fixing antibody titers to Coxiella burnetii phase I (reciprocal titer above 200). With the enzyme-linked immunosorbent assay (ELISA), titers of immunoglobulin G (IgG) to phases I and II of C. burnetii averaged 158,000 and 69,900, respectively, whereas they reached 300 and 3,200 in acute Q fever cases. Similarly, IgA to both phases of C. burnetii and IgM to phase I were consistently higher during chronic than acute Q fever. The serological follow-up of one patient with chronic Q fever over a 4-year period showed a good correlation between the titers of IgG and IgM antibody titers detected by ELISA and indirect fluorescent-antibody test (IFA) to both phases of C. burnetii. Few discrepancies appeared with IgA. Shortly after initiation of antibiotic treatment, a slow and steady decrease of the antibody titers to C. burnetii phases I and II was observed. The complement fixation, IFA, and ELISA tests showed the same type of antibody response. The ELISA proved to be an excellent diagnostic test for chronic Q fever. It distinguished negative from positive reactions clearly, and results were highly reproducible. The reading is objective, and the test is simple to perform and more sensitive than the IFA and complement fixation tests. The ELISA is recommended for serologic evaluation of patients with chronic Q fever.

An enzyme-linked immunosorbent assay (ELISA) was developed to detect immunoglobulin G to Coxiella burnetii phase II. Serum samples from 213 patients who had had Q fever 1 year previously and from 301 blood donors from six localities in Switzerland were tested by ELISA and by indirect fluorescent-antibody (IFA) and complement fixation (CF) tests. The ELISA and the IFA and CF tests detected antibody to C. burnetii in 202 (94.8%), 193 (90.6%), and 166 (77.8%) of the 213 Q fever patients, respectively. With the serum samples from blood donors, the ELISA yielded a higher percentage of positive sera than did the IFA and CF tests. The high specificity of the three tests was confirmed by analyzing paired serum samples from 36 patients suffering from acute pneumonia of viral or bacterial origin. In these cases, the serological results were negative by the three tests, except for three Q-fever cases included as positive control.

Knowledge of the development of different classes of antibody during the course of acute Q fever is important to the clinician for interpreting a patient's serological test results. In the present study, the appearance of antibodies to Coxiella burnetii phases I and II was determined for a period of 1 year. A total of 683 sera from 191 patients with symptomatic Q fever were evaluated by the complement fixation and indirect immunofluorescence (immunoglobulins M and G [IgM, IgG]) tests. These patients had contracted acute Q fever in the fall of 1983 during an epidemic that resulted in 415 serologically confirmed cases of Q fever. As demonstrated by the complement fixation test, antibodies to C. burnetii phase II remained elevated throughout the entire study period, whereas antibodies to phase I were barely detectable. Although the immunofluorescence test was more sensitive than the complement fixation test, the specific anti-IgG response to C. burnetii to phases I and II gave the same general antibody profiles as did the complement fixation test. IgM anti-phase I and II titers appeared earlier but disappeared after 10 to 12 weeks. During this period, anti-phase II antibody levels were generally much higher than anti-phase I antibody levels.