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Background Tick-borne encephalitis (TBE) is an important tick-borne disease in Europe. Detection of the TBE virus (TBEV) in local populations of Ixodes ricinus ticks is the most reliable proof that a given area is at risk for TBE, but this approach is time-consuming and expensive. A cheaper and simpler approach is to use immunology-based methods to screen vertebrate hosts for TBEV-specific antibodies and subsequently test the tick populations at locations with seropositive animals. Results The purpose of the present study was to use goats as sentinel animals to identify new risk areas for TBE in the canton of Valais in Switzerland. A total of 4114 individual goat sera were screened for TBEV-specific antibodies using immunological methods. According to our ELISA assay, 175 goat sera reacted strongly with TBEV antigen, resulting in a seroprevalence rate of 4.3%. The serum neutralization test confirmed that 70 of the 173 ELISA-positive sera had neutralizing antibodies against TBEV. Most of the 26 seropositive goat flocks were detected in the known risk areas in the canton of Valais, with some spread into the connecting valley of Saas and to the east of the town of Brig. One seropositive site was 60 km to the west of the known TBEV-endemic area. At two of the three locations where goats were seropositive, the local tick populations also tested positive for TBEV. Conclusion The combined approach of screening vertebrate hosts for TBEV-specific antibodies followed by testing the local tick population for TBEV allowed us to detect two new TBEV foci in the canton of Valais. The present study showed that goats are useful sentinel animals for the detection of new TBEV risk areas. Electronic supplementary material The online version of this article (doi:10.1186/s12917-017-1136-y) contains supplementary material, which is available to authorized users.

Background Throughout Europe, Ixodes ricinus transmits numerous pathogens. Its widespread distribution is not limited to rural but also includes urbanized areas. To date, comprehensive data on pathogen carrier rates of I. ricinus ticks in urban areas of Switzerland is lacking. Results Ixodes ricinus ticks sampled at 18 (sub-) urban collection sites throughout Switzerland showed carrier rates of 0% for tick-borne encephalitis virus, 18.0% for Borrelia burgdorferi (sensu lato), 2.5% for Borrelia miyamotoi, 13.5% for Rickettsia spp., 1.4% for Anaplasma phagocytophilum, 6.2% for "Candidatus Neoehrlichia mikurensis", and 0.8% for Babesia venatorum (Babesia sp., EU1). Site-specific prevalence at collection sites with n > 45 ticks (n = 9) significantly differed for B. burgdorferi (s.l.), Rickettsia spp., and "Ca. N. mikurensis", but were not related to the habitat type. Three hundred fifty eight out of 1078 I. ricinus ticks (33.2%) tested positive for at least one pathogen. Thereof, about 20% (71/358) were carrying two or three different potentially disease-causing agents. Using next generation sequencing, we could detect true pathogens, tick symbionts and organisms of environmental or human origin in ten selected samples. Conclusions Our data document the presence of pathogens in the (sub-) urban I. ricinus tick population in Switzerland, with carrier rates as high as those in rural regions. Carriage of multiple pathogens was repeatedly observed, demonstrating the risk of acquiring multiple infections as a consequence of a tick bite. Electronic supplementary material The online version of this article (10.1186/s13071-017-2500-2) contains supplementary material, which is available to authorized users.

Coxiella burnetii infection (Q fever) is a widespread zoonosis with low endemicity in Switzerland, therefore no mandatory public report was required. A cluster of initially ten human cases of acute Q fever infections characterized by prolonged fever, asthenia and mild hepatitis occurred in 2012 in the terraced vineyard of Lavaux. Epidemiological investigations based on patients’ interviews and veterinary investigations included environmental sampling as well as Coxiella-specific serological assay and molecular examinations (real-time PCR in vaginal secretions) of suspected sheep. These investigations demonstrated that 43% of sheep carried the bacteria whereas 30% exhibited anti-Coxiella antibodies. Mitigation measures, including limiting human contacts with the flock, hygiene measures, flock vaccination and a public official alert, have permitted the detection of four additional human cases and the avoidance of a much larger outbreak. Since November 2012, mandatory reporting of Q fever to Swiss public health authorities has been reintroduced. A close follow up of human cases will be necessary to identify chronic Q fever.

The most frequent clinical manifestation of borreliosis in Switzerland is erythema migrans, with about 2500 patients each year. Neurological manifestations are rare, mostly hyperalgesic radiculitis (Bannwarth syndrome), aseptic meningitis or cranial nerve involvement. We report the first Swiss patient with meningovasculitis due to neuroborreliosis, with recurrent multiple ischemic strokes in multiple vascular territories. The treatment with ceftriaxone stopped the progression, but the patient is still suffering from severe invalidating cognitive disorders. We also comment on the pathophysiology and review the literature of other clinical cases.

L’infezione da Borrelia burgdorferi (malattia o meglio borreliosi di Lyme) e una spirochetosi trasmessa da zecche ed e endemica, in focolai, in Nord America, Europa e Asia. Questa modalita di infestazione spiega la sua presentazione stagionale. Le manifestazioni cliniche sono a esordio focale, con estensione sistemica; gli organi piu interessati sono la cute, il cuore, il sistema muscoloscheletrico, il sistema nervoso, spesso con interessamento sequenziale. L’infezione precoce, evidenziata dall’eritema migrante, e seguita da uno stadio di disseminazione nelle settimane successive; i sintomi intermittenti possono persistere e provocare manifestazioni tardive osservate in media entro l’anno. In un paziente infetto tutti gli stadi possono essere presenti oppure puo non comparirne nessuno. I sintomi neurologici si verificano nello stadio piu precoce (radicolonevrite, neuropatie craniche, meningite, sintomi cerebrali minori, miosite) o tardivo (encefalomielite progressiva, encefalopatia, neuropatia assonale). Tra i casi europei e americani esistono numerose differenze, dovute alle specie genotipiche diverse di Borrelia burgdorferi nei due continenti. In effetti, la linfadenosi cutanea benigna, l’acrodermatite cronica atrofica e l’encefalomielite progressiva sono osservate quasi unicamente in Europa, perche Borrelia afzelii e garinii, non osservate nel Nord America, ne sono la probabile causa. La sierologia e la PCR (polymerase chain reaction) sono attualmente i mezzi a disposizione, che messi in relazione con i sintomi clinici, permettono di porre la diagnosi di borreliosi di Lyme. Esistono diversi antibiotici efficaci, che permettono un trattamento dell’infezione precoce; le manifestazioni tardive rispondono parzialmente alla terapia. Nessun vaccino e attualmente disponibile.

Coxiella burnetii and members of the genus Rickettsia are obligate intracellular bacteria. Since cultivation of these organisms requires dedicated techniques, their diagnosis usually relies on serological or molecular biology methods. Immunofluorescence is considered the gold standard to detect antibody-reactivity towards these organisms. Here, we assessed the performance of a new automated epifluorescence immunoassay (InoDiag) to detect IgM and IgG against C. burnetii, Rickettsia typhi and Rickettsia conorii. Samples were tested with the InoDiag assay. A total of 213 sera were tested, of which 63 samples from Q fever, 20 from spotted fever rickettsiosis, 6 from murine typhus and 124 controls. InoDiag results were compared to micro-immunofluorescence. For acute Q fever, the sensitivity of phase 2 IgG was only of 30% with a cutoff of 1 arbitrary unit (AU). In patients with acute Q fever with positive IF IgM, sensitivity reached 83% with the same cutoff. Sensitivity for chronic Q fever was 100% whereas sensitivity for past Q fever was 65%. Sensitivity for spotted Mediterranean fever and murine typhus were 91% and 100%, respectively. Both assays exhibited a good specificity in control groups, ranging from 79% in sera from patients with unrelated diseases or EBV positivity to 100% in sera from healthy patients. In conclusion, the InoDiag assay exhibits an excellent performance for the diagnosis of chronic Q fever but a very low IgG sensitivity for acute Q fever likely due to low reactivity of phase 2 antigens present on the glass slide. This defect is partially compensated by the detection of IgM. Because it exhibits a good negative predictive value, the InoDiag assay is valuable to rule out a chronic Q fever. For the diagnosis of rickettsial diseases, the sensitivity of the InoDiag method is similar to conventional immunofluorescence.

Background Different tick-borne infections can cause an acute febrile illness. The study objectives were to investigate the clinical manifestations and diagnosis of infections among patients who presented with fever after a tick-bite, and to detect newly described pathogens, including Ehrlichia, Babesia and Rickettsia helvetica, in North-Eastern Switzerland. Patients and methods : We studied 75 patients (41 male, 34 female, median age 38 years, among them 10 children) who had fever within 3 weeks after a tick-bite. Paired sera were tested for antibodies to Borrelia burgdorferi, tick-borne encephalitis virus, Anaplasma (Ehrlichia) phagocytophila, Babesia microti, B. divergens, and Rickettsia helvetica. In addition, microscopy and polymerase chain reaction was used to detect Ehrlichia. Clinical data were obtained at baseline and at 1 and 2 year follow-up. Results Tick-borne infections were confirmed or possible in 36 (48 %) patients: 7 (9 %) Erythema migrans, 6 (8 %) other specific manifestations of Lyme borreliosis, 6 (8 %) Lyme borreliosis presenting as non-specific febrile illness, 8 (11 %) tick-borne encephalitis, 7 (10 %) granulocytic ehrlichiosis, 1 B. microti infection in a traveler from the US and 6 (8 %) dual infections. In 8 (11 %) patients serological findings were suggesting possible acute or past R. helvetica infection. Conclusion Among patients with fever after a tick-bite, Lyme borreliosis was most frequently found. There was no evidence for babesiosis among the resident population. Serologic data suggest that human granulocytic ehrlichiosis and R. helvetica infections may be endemic in Switzerland. Among 50 % of the patients no tick-borne infections could be diagnosed.

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.

Resume Avant de chercher a combattre un parasite, il est indispensable de bien le connaitre. Plus de 800 especes de tiques ont ete recensees dans le monde entier. Elles parasitent des hotes tres varies et ont des modes de vie fort differents d’une espece de tique a l’autre. Il est extremement difficile de controler ces ectoparasites et les maladies qu’elles transmettent. A l’heure actuelle le controle de l’infestation des animaux domestiques repose essentiellement sur les acaricides, sous forme de bains, de sprays, de shampooing, de colliers ou de boules de ouate impregnes… Des croisements genetiques de differentes races de bovins permettent d’accroitre la resistance naturelle de ces animaux contre les tiques. Des vaccins anti-tiques sont actuellement commercialises en Australie et a Cuba pour le betail contre Boophilus microplus et des recherches se poursuivent pour le developpement d’autres vaccins anti-tiques. Dans le futur, des luttes specifiques integrees, prenant en compte les caracteristiques des especes de tiques impliquees, devraient ameliorer le controle de ces ectoparasites.

Resume Cette etude presente de maniere nouvelle et detaillee la complexite de la reponse immunologique a une borreliose de Lyme, telle que nous avons pu l'observer avec les immunoblots que nous avons mis au point. Elle s'efforce egalement de signaler les anticorps les plus caracteristiques de chaque stade de la maladie et illustre l'importance de l'echange d'information entre biologistes et cliniciens. Cette coordination permet de definir les meilleures strategies d'investigation et d'ameliorer la fiabilite des examens de laboratoire tout en reduisant leurs couts.

OBJECTIVE: To evaluate by species-specific immunoblots the association of Borrelia burgdorferi sensu stricto, B. garinii and B. afzelii with neuroborreliosis in Switzerland. METHODS: Borrelia strains isolated from the cerebrospinal fluid (CSF) of three children with neuroborreliosis were typed by phenotypic and genotypic analysis. The serologic reactions (IgG) of these three patients as well as those of 28 patients, including one of these three children, with confirmed neuroborreliosis were characterized and scored by immunoblots on the three individual Borrelia species antigens. Twenty patients with typical erythema migrans served as a control group. RESULTS: Phenotypic and genotypic analysis confirmed that all three CSF isolates were B. garinii. In the 28 patients with neuroborreliosis, the comparatively strongest reactions were as follows: 18 to B. garinii, three to B. burgdorferi sensu stricto and two to B. afzelii; five were inconclusive. In the control group (erythema migrans), the comparatively strongest reactions were as follows: six B. garinii, one to B. burgdorferi sensu stricto and five to B. afzelii; eight were indeterminate. CONCLUSIONS: Typing of these three CSF isolates and characterization by immunoblots of the antibody reactions of patients with neuroborreliosis give additional evidence of the association of B. garinii and neuroborreliosis. Our serologic results suggest that B. burgdorferi sensu stricto and B. afzelii are also responsible for some neuroborreliosis cases in Switzerland. Our immunoblots and the scoring system proved particularly useful for the serologic typing of patients with late Lyme borreliosis.

Summary QUESTION UNDER STUDY: To determine the incidence and determinants of tick related diseases in Switzerland, such as tick bites and Lyme borreliosis in primary care and tick borne encephalitis. METHODS: Analysis of the Swiss data collected by mandatory and facultative surveillance systems for the reporting period of 2008 to 2011. RESULTS: Tick related diseases in Switzerland are common. About 17,000 to 23,000 estimated cases of tick bites lead to a consultation (yearly incidence 254 per 100,000 inhabitants); about 7,000 to 12,000 estimated cases of Lyme borreliosis (yearly incidence 131 per 100,000 inhabitants) and 98 to 172 cases of tick borne encephalitis occur each year (yearly incidence 1.6 per 100,000 inhabitants). The most affected area is the north-eastern part of Switzerland. Whereas cases of tick borne encephalitis are restricted to local endemic areas, cases of Lyme borreliosis and tick bites are spread all over Switzerland. CONCLUSIONS: Tick related diseases are frequent and widespread in Switzerland. They are leading to a considerable usage of the health care system. Thus, tick bite prevention and vaccination against tick borne encephalitis are essential. However, long term follow-up cohort studies with reasonably large study populations after tick bite would be required to elucidate the risk of developing a tick borne disease.

We describe two cases of Q fever in previously healthy women presenting with fever of unknown origin. The diagnosis was made after several days of investigations. Symptoms and signs of acute or chronic Coxiella burnetii infection are protean and non-specific. Q fever should be included in the differential diagnosis of fever of unknown origin and appropriate serologic studies should be done. We review the clinical presentation of Q fever. Use of serology for the diagnosis and the follow-up is discussed.

Tick-borne encephalitis (TBE, in French MEVE) is a viral disease usually transmitted to man through an infected tick belonging to the genus Ixodes. Every year about 135 cases of TBE are reported to the Swiss Federal Office of Public Health. Emerging in north-east of Switzerland in the seventies, endemic areas have been slowly spreading. Actually 18 cantons out of 26 have been reached. In the western part of Switzerland, spared until not long ago, new endemic foci are spreading toward the lake of Geneva and recently in Valais. The precise exposure history of human cases, as well as the evaluation of collected tick samples allow the detection of endemic foci which are discontinuous and of variable size. New and more efficient methods are needed to monitor the changing epidemiology of TBE.

The Chlamydiales order includes the Chlamydiaceae, Parachlamydiaceae, Waddliaceae, Simkaniaceae, Criblamydiaceae, Rhabdochlamydiaceae, Clavichlamydiaceae, and Piscichlamydiaceae families. Members of the Chlamydiales order are obligate intracellular bacteria that replicate within eukaryotic cells of different origins including humans, animals, and amoebae. Many of these bacteria are pathogens or emerging pathogens of both humans and animals, but their true diversity is largely underestimated, and their ecology remains to be investigated. Considering their potential threat on human health, it is important to expand our knowledge on the diversity of Chlamydiae, but also to define the host range colonized by these bacteria. Thus, using a new pan-Chlamydiales PCR, we analyzed the prevalence of Chlamydiales DNA in ticks and fleas, which are important vectors of several viral and bacterial infectious diseases. To conduct this study, 1340 Ixodes ricinus ticks prepared in 192 pools were collected in Switzerland and 55 other ticks belonging to different tick species and 97 fleas belonging to different flea species were harvested in Algeria. In Switzerland, the prevalence of Chlamydiales DNA in the 192 pools was equal to 28.1% (54/192) which represents an estimated prevalence in the 1340 individual ticks of between 4.0% and 28.4%. The pan-Chlamydiales qPCR was positive for 45% (25/55) of tick samples collected in Algeria. The sequencing of the positive qPCR amplicons revealed a high diversity of Chlamydiales species. Most of them belonged to the Rhabdochlamydiaceae and Parachlamydiaceae families. Thus, ticks may carry Chlamydiales and should thus be considered as possible vectors for Chlamydiales propagation to both humans and animals.

Summary Nine strains of Borrelia burgdorferi isolated from ticks in the canton of Valais (Switzerland) were characterized genotypically by determining restriction fragment length polymorphisms (RFLP) and plasmid profiles. The strains were also compared with respect to presence and electrophoretic mobility of the outer surface proteins OspA and OspB, and immunoreactivity of OspA and a 12 kD antigen. By both approaches, three different patterns were observed resulting in identical grouping of the strains. However, RFLP's allowed determination of relationships among strains within a group and have shown that geographic distribution does not correlate with genotype.

Among the 20 tick species described in Switzerland, Ixodes ricinus, the most frequent one, is implicated in the transmission of pathogenic agents. Lyme borreliosis and tick-borne encephalitis (TBE) are the major tick-borne diseases transmitted to human. Presently 5 Borrelia species, belonging to the group Borrelia burgdorferi, are recognized as human pathogens. The risks of infection depend on the stage of the vector, the multiple hosts, the pathogenic agent, as well as human behavior in nature. The detection of other pathogenic agents in ticks: Anaplasma, Babesia and Rickettsia predispose to infections or co-infections. Results of sero-epidemiologic studies suggest human infections. Active surveillance by physicians is necessary and clinical studies are required to evaluate the importance of these infections in Switzerland.

The most frequent clinical manifestation of borreliosis in Switzerland is erythema migrans, with about 2500 patients each year. Neurological manifestations are rare, mostly hyperalgesic radiculitis (Bannwarth syndrome), aseptic meningitis or cranial nerve involvement. We report the first Swiss patient with meningovasculitis due to neuroborreliosis, with recurrent multiple ischemic strokes in multiple vascular territories. The treatment with ceftriaxone stopped the progression, but the patient is still suffering from severe invalidating cognitive disorders. We also comment on the pathophysiology and review the literature of other clinical cases.

Coxiella burnetii and Brucella abortus are two intracellular bacteria implicated in zoonotic miscarriage. In the present study, C. burnetii and B. abortus seroprevalence was compared among women from London with and without miscarriage. Coxiella burnetii seroprevalence was high (4.6%, 95% CI 2.8–7.1) despite the rare apparent exposure of this urban population. Only two patients exhibited anti- B. abortus antibodies. As a result of the risk of chronic Q fever with endocarditis and/or hepatitis, the mode of Coxiella burnetii infection in this population merits further investigation.

BACKGROUND: HSV-1 and HSV-2 cause CNS infections of dissimilar clinico-pathological characteristics with prognostic and therapeutic implications. OBJECTIVES: To validate a type-specific real-time PCR that uses MGB/LNA Taqman probes and to review the virologico-clinical data of 25 eligible patients with non-neonatal CNS infections. RESULTS: This real-time PCR was evaluated against conventional PCR (26 CSF and 20 quality controls), and LightCycler assay (51 mucocutaneous, 8 CSF and 32 quality controls) and culture/immunofluorescence (75 mucocutaneous) to assess typing with independent methods. Taqman real-time PCR detected 240 HSV genomes per ml CSF, a level appropriate for the management of patients, and provided unambiguous typing for the 104 positive (62 HSV-1 and 42 HSV-2) out the 160 independent clinical samples tested. HSV type diagnosed by Taqman real-time PCR predicted final diagnosis (meningitis versus encephalitis/meningoencephalitis, p

Lyme disease is the most common tick-borne disease in Europe and in the United States. In comparison to dermatological, neurological and rheumatological manifestations, heart disease is quite rare. Atrioventricular heart block is nevertheless the most frequent cardiological manifestation. We hereby report the case of a patient with high degree heart block due to Lyme disease. We focus on the electrocardiographical evolution during antibiotic therapy, as well as on microbiological and diagnostic aspects. Lyme disease is a rare cause of conduction disturbances but it is treatable and potentially reversible.

The aim of this study was to define the risk of developing Lyme borreliosis after a tick bite. A survey was conducted from 1993 to 1995 in the western part of Switzerland in a group of patients who presented for treatment of a recent tick bite. Only patients with negative serological tests (enzyme-linked fluorescent assay screening test, and IgG and IgM immunoblots) at the first consultation and for whom a second blood sample was available 2 months later were included in the study. Of the 376 patients included, 266 had no clinical manifestation (group 1) and 110 had a small local cutaneous reaction (

BACKGROUND: Although European Borrelia burgdorferi sensu lato isolates have been divided into five genospecies, specific tools for the serotype characterization of only three genospecies are available. Monoclonals antibodies (mAbs) H3TS, D6 and I17.3 identify B. burgdorferi sensu stricto (ss.), B. garinii and B. afzelii respectively, but no mAbs are available to identify B. valaisiana. In the same way, specific primers exist to amplify the OspA gene of B. burgdorferi ss. B. garinii and B. afzelii. The aim of the study was to develop species-specific mAb and PCR primers for the phenotypic and genetic identification of B. valaisiana. RESULTS: This study describes a mAb that targets OspA of B. valaisiana and primers targeting the OspA gene of this species. As the monoclonal antibody A116k did not react with strains NE231, M7, M53 and Frank and no amplification was observed with strains NE231, M7 and M53, the existence of two subgroups among European B. valaisiana species was confirmed. CONCLUSIONS: The association of both monoclonal antibody A116k and primers Bval 1F and Bval 1R allows to specific identification of the B. valaisiana isolates belonging to subgroup 1., Journal Article, Research Support, Non-U.S. Gov't, SCOPUS: ar.j, info:eu-repo/semantics/published

Different tick-borne infections can cause an acute febrile illness. The study objectives were to investigate the clinical manifestations and diagnosis of infections among patients who presented with fever after a tick-bite, and to detect newly described pathogens, including Ehrlichia, Babesia and Rickettsia helvetica, in North-Eastern Switzerland.: We studied 75 patients (41 male, 34 female, median age 38 years, among them 10 children) who had fever within 3 weeks after a tick-bite. Paired sera were tested for antibodies to Borrelia burgdorferi, tick-borne encephalitis virus, Anaplasma (Ehrlichia) phagocytophila, Babesia microti, B. divergens, and Rickettsia helvetica. In addition, microscopy and polymerase chain reaction was used to detect Ehrlichia. Clinical data were obtained at baseline and at 1 and 2 year follow-up.Tick-borne infections were confirmed or possible in 36 (48 %) patients: 7 (9 %) Erythema migrans, 6 (8 %) other specific manifestations of Lyme borreliosis, 6 (8 %) Lyme borreliosis presenting as non-specific febrile illness, 8 (11 %) tick-borne encephalitis, 7 (10 %) granulocytic ehrlichiosis, 1 B. microti infection in a traveler from the US and 6 (8 %) dual infections. In 8 (11 %) patients serological findings were suggesting possible acute or past R. helvetica infection.Among patients with fever after a tick-bite, Lyme borreliosis was most frequently found. There was no evidence for babesiosis among the resident population. Serologic data suggest that human granulocytic ehrlichiosis and R. helvetica infections may be endemic in Switzerland. Among 50 % of the patients no tick-borne infections could be diagnosed.

OBJECTIVE: To test the performances of new Borrelia garinii immunoblots specific for Borrelia burgdorferi sensu lato with a selected panel of sera from patients with various clinical presentations of Lyme borreliosis. METHODS: In order to establish the sensitivity and the specificity of these immunoblots, we tested serum samples obtained from patients with early- and late-stage Lyme disease (erythema migrans n=35, neuroborreliosis n=61, acrodermatitis chronica atrophicans (ACA) n=27 and arthritis n=41), from patients with diagnoses and laboratory findings associated with serologic cross-reactivity to Lyme disease (syphilis n=12, Epstein-Barr infection n=9, autoimmune markers n=29) and from blood donors residing in regions of low and medium endemicity (n=80, n=100). RESULTS: The combined sensitivity (IgG and IgM) of the tests was 90% for patients with erythema migrans, 92% for neuroborreliosis, 96% for ACA and 100% for Lyme arthritis. The specificity of the IgG immunoblot was 94%, and that of the IgM immunoblot was 97%, taking into account the prevalence of borrelia antibodies in the overall population. Interpretation of these immunoblots is based on scores allocated to different specific borrelia antigens. CONCLUSIONS: The Western blot technology is extremely useful in dissecting the immune response to borrelia infections, which develops gradually over a period of weeks to years and which involves the appearance of IgM and IgG antibodies directed against a number of borrelia-associated proteins.

In the course of respiratory infections, the efficacy of microbiologic diagnosis has increased years after years, in term of specificity, sensitivity and rapidity. New pathogenic agents have been described such as: Legionella pneumophila, Chlamydia pneumoniae, Hantavirus. Some viruses have been well characterized as responsible for seasonal outbreaks using rapid tools for identification. Needs for efficient diagnostic tools became more obvious when specific antiviral drugs appeared on the market. So technologic developments improved the efficacy of microbiologic diagnosis and anticipate a better specificity as well as sensitivity with the help of molecular biology. Respiratory syncytial virus is one of the major infectious agents found in respiratory infections in young children and newborns. On the whole it was detected in more than one third of pediatric nasopharyngeal aspirations received in our laboratory and more than 50% during the peak of the winter epidemics. The method of direct antigen detection by immunofluorescence with the help of monoclonal antibodies allowed us to establish an incidence curve of these recurrent outbreaks, beginning in December to stop usually by the end of April. During this same period, influenza A virus, seldom influenza B virus, were detected in many nasopharyngeal specimens. Other viruses, parainfluenza 1 to 3 and Adenovirus, were irregularly detected all along the year. In the great majority of nasopharyngeal aspirations with a positive virus detection, one virus only was observed. Antigen detection methods were also developed for some bacteria such as Chlamydia pneumoniae, Legionella pneumophila. Although serology is not frequently used by pediatricians, it is still necessary for the diagnosis of Mycoplasma pneumoniae infections. A direct antigen detection test is now available, but its sensitivity needs to be evaluated. On the other hand serologic diagnosis may be extremely useful when long lasting or treatment resistant respiratory infections occur. Seroconversion or four-fold increasing titers to one pathogen may be observed when a second serum sample is tested together with the first serum of this patient. The diagnostic yield will be all the more efficient that time between both samples is long. Molecular biology techniques will significantly change the way to investigate an infection. Presently these methods are used in research laboratories, but automated technologies will facilitate routine laboratory workload. Screening methods using multiplex PCR are also promising.

Borrelia garinii is one of the three major Borreliae responsible for Lyme borreliosis in Europe. We have characterized a protein of B. garinii (VS102) and a genomic fragment from the gene encoding this protein was cloned. The DNA sequence of the fragment showed high homology with a known gene of B. burgdorferi sensu stricto. The protein encoded by this gene in B. burgdorferi sensu stricto is a phosphocarrier protein (histidine-containing protein). A mutation T to G polymorphism at codon 57 was found to be specific to B. garinii. A PCR-based approach that allows the rapid detection of this mutation made it possible to specifically discriminate B. garinii from other B. burgdorferi genospecies with high sensitivity and specificity.

Objective To evaluate the range of long term vascular manifestations of Coxiella burnetii infection. Design Cohort study in Switzerland of people affected in 1983 by the largest reported outbreak of Q fever and who were followed up 12 years later. Follow up information about possible vascular disease and endocarditis was obtained through a mailed questionnaire and death certificates. Setting Val de Bagnes, a rural Alpine valley in Switzerland. Participants 2044 (87%) of 2355 people who had serum testing for Coxiella burnetii infection in 1983: 1247 were classed as not having been infected, 411 were classed as having been acutely infected, and 386 were classed as having been infected before 1983. Main outcome measures Relative risk controlled for age and sex and 12 year risk of vascular diseases and endocarditis among infected participants as compared with those who had never been infected. Results The 12 year risk of endocarditis or venous thromboembolic disease was not increased among those who had been acutely infected. The 12 year risk of arterial disease was significantly higher among those who had been acutely infected (7%) as compared with those who had never been infected (4%) (relative risk 2.2, 95% confidence interval 1.4 to 3.6). Specifically, there was an increased risk of developing a cerebrovascular accident (relative risk 3.7, 1.6 to 8.4) and cardiac ischaemia (relative risk 1.9, 1.04 to 3.4). 12 year mortality was significantly higher among the 411 people who had been acutely infected in 1983 (9.7%; age adjusted relative risk 1.8, 1.2 to 2.6) when compared with the 1247 participants who had remained serologically negative in 1983 (7.0%). Conclusions Coxiella burnetii infection may cause long term complications including vascular disease.

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.

The purpose of this study was to analyse the clinical and serological follow-up in 21 patients with Q fever endocarditis in Switzerland from 1981 to 1993.Criteria for Q fever endocarditis were the following: Coxiella burnetii phase I IgG1 : 2560 and IgA1 : 20 by indirect immunofluorescence. Methods to confirm the diagnosis include immunohistochemical demonstration of C. burnetii by microscopy in valvular material (1 case) and inoculation of this material in experimental animals (10 cases). Information on clinical course of the disease, laboratory abnormalities and treatment were obtained by chart review and a questionnaire sent to physicians who requested the serological tests for Q fever.The average age of the patients was 47 years (15 men and 6 women). 64% of patients had a history of environmental exposure to C. burnetii. The median time of symptomatology before diagnosis was 5 months (1-108). 19/21 patients had valvular lesions, and 2/21 vascular Dacron prosthesis. Most patients presented with fever (18/21), congestive cardiac failure (14/21), weight loss (12/21), anemia (6/19), or thrombocytopenia (6/19). All the patients required antibiotic treatment. Cardiac surgery was performed in 15/21 patients. For 10 patients the geometric mean serological follow-up included at least titers at time of diagnosis (IgG anti-phase I antibodies 1 : 27024, IgA anti-phase I antibodies 1 : 685), at the end of therapy (IgG anti-phase I antibodies 1 : 2941, IgA anti-phase I antibodies 1 : 153) and 6 months after the end of therapy (IgG anti-phase I antibodies 1 : 368, IgA anti-phase I antibodies 1 : 40). The fall in anti-phase I titers was significant. During the clinical and serological outcome (median of 60 months and 69 months respectively) there was no recurrence of endocarditis and antibody titers to C. burnetii phase I remained low. Two patients died during the observation period, one from lung cancer, while the cause of death in the other was unknown.Serology is the key to Q fever diagnosis. The duration of treatment, and the values to be used to establish cure of endocarditis, are not clearly defined. During the clinical and serological outcome (median of 60 months and 69 months respectively) there was no recurrence of endocarditis and antibody titers to C. burnetii phase I remained low.

The diversity and the distribution of tick species and their infection rates by the pathogenic micro-organism Borrelia burgdorferi sensu lato, the etiologic agent of Lyme borreliosis, and Rickettsia sp., were studied in Canton Ticino (the southern part of Switzerland). Ticks specimens collected from animals and humans were classified and analysed for the presence of both pathogens. In particular, PCR analysis was performed for the detection of Borrelia spirochetes in Ixodes ricinus and Ixodes hexagonus, and the hemolymph test was done on Rhipicephalus sanguineus for the detection of Rickettsia sp. PCR assays, performed on 424 of the 989 collected ticks, revealed a low rate of infection (around 2%) of both vectors I. ricinus and I. hexagonus by B. burgdorferi sensu lato. These results are in agreement with the modest number of Lyme borreliosis cases yearly recorded in Ticino. Further, through analysis of DNA sequences, the strains carried by the infected ticks were identified as belonging to the genomic group VS116. The widespread finding of the Mediterranean species Rhipicephalus sanguineus in different locations from July 1994 to October 1995 demonstrates its establishment in Ticino. Of the 210 specimens collected, 70 were analysed and one was infected by Rickettsia sp.

A total of 825 adult ticks (727 Ixodes ricinus, 72 Dermacentor marginatus and 26 Haemaphysalis punctata) was collected from vegetation in Valais (Switzerland) in 1987 to 1992. They were examined for the presence of Borrelia burgdorferi sensu lato, the etiologic agent of Lyme borreliosis. B. burgdorferi sensu lato was detected by indirect immunofluorescence assay, dark field microscopy and/or culture in 221 out of 727 I. ricinus (30.4%) and none in the other two species. From these 221 infected ticks we obtained 50 isolates. Indirect immunofluorescence assay and culture were used for all ticks but dark field examination has also been performed and compared to the two above mentioned methods for 231 I. ricinus. Indirect immunofluorescence assay and culture were used for all ticks but dark field examination has also been performed and compared to the two above mentioned methods for 231 I. ricinus. Indirect immunofluorescence was found the most efficient method for the detection of Borrelia in ticks with 54 positive out of 231, followed by dark field examination with 35 positive and culture with 12 isolates. We found no site free of Borrelia where I. ricinus is present. The rate of infection varied from 9.7 to 47.5%, as detected by the addition of the three methods. Typing of the 50 isolates revealed also a nonhomogeneous distribution of the Borrelia species. Based on the electrophoretic mobility of the OspA and B and immunostaining with species specific monoclonal antibodies (H3TS for B. burgdorferi sensu stricto, D6 for B. garinii and J8.3 for B. afzelii) 4 groups could be observed. Half of the isolates (n = 26) were typed as B. burgdorferi sensu stricto, 19 as B. garinii, 3 as B. afzelii and 2 as group VS116. This forth group formed of two isolates from one location is genetically distinct from the 3 former species described in Europe so far. The Borreliae of this group are unreactive with any of the three monoclonal antibodies used.

Isolation of Borrelia burgdorferi from the CSF is relatively rare. The present report describes the first three isolations in Switzerland. Clinically, our first observation confirmed the frequent association of B. burgdorferi with peripheral facial paresis in children. The other two cases illustrate the variety of symptoms in neuro-borreliosis. In the first case the culture was positive after 6 weeks. The results of serologic tests (indirect immunofluorescence and ELISA) for detection of antibodies against B. burgdorferi were negative or non-significant in this child's serum. On the other hand, specific antibodies (IgG) were detected in the serum by western blot. Culture of the second CSF already showed Borrelia growth after 10 days. Immunofluorescence revealed high antibody titers (1/256) against B. burgdorferi in this patient's serum. IgG showed a weakly positive reaction in western blot. The reliability of this result was confirmed by isolation of Borrelia. In neither of the two CSF could intrathecal synthesis of specific antibodies be demonstrated. In the third case, however, immunofluorescence showed IgG antibody titers of 1/128 in the CSF and 1/512 in serum. Intrathecal synthesis of specific antibodies was demonstrated with an index of 13.4 (norm2). Western blot confirmed the specificity of the reactions observed with the serum and CSF IgG. Culture of CSF produced significant growth of Borrelia within 7 days. Protein profile and reactions with poly- and monoclonal antibodies confirmed that the three strains belonged to B. burgdorferi.(ABSTRACT TRUNCATED AT 250 WORDS)

The scope of current diagnostic methods for Q fever endocarditis includes serology, direct demonstration of Coxiella burnetii in the resected heart valve tissue, and animal inoculation studies. Illustrated by a clinical case report, the different methods are presented and discussed. Serology represents the primary method, using the techniques of complement fixation, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA). The latter two techniques allow the detection of immunoglobulins G, M, and A to the phase I and II antigens of C. burnetii. After cardiac surgery, we visualized C. burnetii on smears and specifically stained it on histologic sections of the resected heart valve by light and electron microscopic immunohistochemistry. In addition, seroconversion in animals after inoculation with valve specimens confirmed the presence of C. burnetii in the heart valve. The antibody titers determined by ELISA correlated well with the patient's clinical course during the treatment period. Therefore it is suggested that its usefulness for monitoring the efficacy of antimicrobial agents in patients with Q fever endocarditis should be further evaluated.

A total of 23 isolates of Borrelia burgdorferi were characterized by SDS-PAGE and immunoblot analysis. One isolate came from the CSF of a Lyme neuro-borreliosis patient in Valais (Switzerland) and 22 were tick isolates (2 from I. dammini of Shelter Island, USA and 20 from I. ricinus of Valais, Switzerland). Based on the electrophoretic mobility of outer surface proteins (OspA and OspB), four groups of B. burgdorferi could be defined. Group I isolates possess an OspA of 31 KD and an OspB of 34 KD. The group II isolate showed an OspA of 32 KD and OspB of 35 KD. Group III isolates have a 33 KD OspA and group IV a 33.5 KD OspA. This classification was confirmed by the reactivity of a monoclonal antibody (D6) to a 12 KD antigen that was recognized in group III only. A Lyme patient's serum showed a 2-band pattern (10 and 13 KD) for group I and a one-band pattern (12 KD) for the other 3 groups. Therefore OspA, OspB and other proteins of low molecular weight (10, 12, and 13 KD) seem to be important keys for the classification of B. burgdorferi isolates. This typing system correlates with genetic analysis.

Fifty Borellia burgdorferi strains isolated from humans and ticks in Europe and the United States were analyzed by multilocus enzyme electrophoresis. Eleven genetic loci were characterized on the basis of the electrophoretic mobilities of their products. Ten loci were polymorphic. The average number of alleles per locus was 5.9, with a mean genetic diversity of 0.673 among electrophoretic types (ETs). The strains were grouped into 35 ETs constituting three main divisions (I, II, and III) separated at a genetic distance greater than 0.75. Divisions I, II, and III contained 13, 6, and 16 ETs, respectively. These findings, together with previous data from DNA hybridization and restriction enzyme analysis of rRNA genes, suggest that divisions I, II, and III may represent three distinct genomic species. All three divisions contained human clinical ETs. However, in division I, which includes the ET of the type strain of B. burgdorferi, the human pathogenic ETs constituted a single clone. The ETs of division I were from west-central Europe and the United States, whereas divisions II and III contained ETs from west-central and northern Europe but not from the United States. Finally, our data show that the genetic structure of B. burgdorferi populations is clonal.

The clinical diagnosis of Lyme borreliosis is easily established in the initial phase of the illness. Erythema chronicum migrans is pathognomonic of this infection. However, during further complications (neurological, cardiac or articular) the diagnosis is chiefly based on laboratory results. The detection of specific antibodies to B. burgdorferi is one of the effective means of confirming the diagnosis. Culture or isolation of this bacteria is not routinely performed due to its very low yield. In this article a case of serologically proved neuro-borreliosis is described and the methods of diagnosis are discussed. Intrathecal synthesis of antibodies observed in the cerebrospinal fluid of this patient and the specificity of serologic tests confirmed by Western-blot provide the diagnosis. However, the absence of locally synthesized antibodies in the cerebrospinal fluid is not sufficient to rule out neuro-borreliosis.

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.

The diagnosis of Q fever endocarditis cannot be made by bacterial cultures and necessitates serological identification of specific antibodies to Coxiella burnetii which stimulates mainly the production of anti-phase II antibodies during the acute disease, but primarily anti-phase I antibodies in endocarditis. Indirect microimmunofluorescence allows rapid detection of specific IgA, IgG and IgM. The results of serological analyses of 191 acute cases of Q fever were compared with those of 8 cases of Coxiella burnetii endocarditis. All sera were evaluated by complement fixation and microimmunofluorescence tests. The highest titre differences between primary Q fever and Q fever endocarditis were observed with anti-phase I IgA and IgG antibodies measured by microimmunofluorescence followed by anti-phase I antibodies measured by complement fixation tests. Antiphase I IgG and IgM titres were consistently higher than anti-phase II titres in endocarditis. The reverse is true in acute Q fever. In addition, anti-phase I IgA appeared to be diagnostic for Coxiella burnetii endocarditis. Accordingly we recommend the testing of these specific IgA, IgG, and IgM by microimmunofluorescence in cases of culture-negative endocarditis. These tests could also prove useful for following the development of Coxiella burnetii endocarditis in patients under treatment.

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.

A comparison was made of the performance of the commonly used complement fixation test and the more recently developed indirect immunofluorescence test in the early diagnosis of Q fever. The 303 sera tested were from 181 patients who contracted Q fever during an outbreak in Switzerland in 1983. Specific IgM antibodies were detected by the immunofluorescence test in 53 % and 89 % of sera obtained during the first and second week respectively after onset of illness. With the complement fixation test, the diagnosis could not be made until the second week of illness. The immunofluorescence test proved to be superior to the complement fixation test in the early detection of Q fever. Not only was it more specific but also faster and simpler to perform, permitting an earlier diagnosis on the basis of results obtained with a single serum specimen.

In current practice, only serology, by the titration of anti-Coxiella burnetii antibodies, permits the diagnosis of acute or chronic Q fever. New serological techniques have also made it possible to more accurately determine the number of people in a community who have been affected by this infectious agent. We have thus been able to compare the results of 1437 specimens from the whole of Switzerland with 4009 taken in the Valais. The analysis of these sera by indirect microimmunofluorescence now shows that Q fever is much more prevalent than was generally estimated. However, these results vary greatly from one area and another, between 6.7% and 31.7% depending on the locality. Normally the lowest percentages were found in the big cities (from 9.5% to 12.2% in Berne, Geneva and Zurich) whilst the highest incidences were found in mountain regions (from 23.6% to 31.7% in the Tessin, Uri or the Valais). Regarding sex distribution, it was found that the incidence of Q fever is on average slightly lower (between 3% and 5%) in women than in men. It was also noted that the percentage of people having anti-Coxiella burnetii antibodies increases with age. The high incidence of Q fever in Switzerland is also of concern because of a severe and chronic form of this infection: endocarditis.

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.