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  • 1
    ISSN: 1432-1440
    Keywords: Lyme arthritis ; Lyme disease ; Tickborne spirochetosis ; Borrelia burgdorferi ; Stomoxys calcitrans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The clinical manifestations, serological data, and radiographic findings of ten cases of Lyme arthritis in Germany are summarized. Qualitative assessment shows that the characteristics of the disease in Germany do not differ fundamentally from those reported in the USA. However, since a serological test for antibodies to the causative spirochete is now available, the great variety of the clinical features of Lyme arthritis can be described more precisely. The cases of chronic Lyme arthritis without prior erythma chronicum migrans, hitherto the most important diagnostic hallmark of the disease, may have been underestimated. One of the cases reported provides evidence that the disease was transmitted via a fly bite. Radiographic abnormalities consisting of marked juxta-articular osteoporosis and osseous erosions were found in two patients with chronic arthritis. Three patients were treated with high-dose intravenous penicillin, two did not respond to the therapy.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A recombinant immunoblot was developed for detection of IgM and IgG antibodies in patients with Lyme borreliosis. The recombinant antigens were the chromosomal-encodedBorrelia burgdorferi proteins p100, the flagellin and an internal flagellin fragment thereof as well as the plasmid-encoded outer surface proteins A (OspA) and C (OspC). A panel of 144 sera from patients with Lyme borreliosis (erythema migrans,n = 31; neuroborreliosis state II,n = 60; Lyme arthritis,n = 24 and acrodermatitis chronica atrophicans,n =19) have been investigated and the results have been compared to the immunofluorescence absorption test (IFA-ABS) and to two different enzyme-linked immunosorbent assays [the flagellin ELISA and a newly developed ELISA (OGP-ELISA)]. The two ELISAs were comparable in sensitivity, whereas the IFA-ABS was less sensitive for IgM antibody but equally sensitive for IgG antibody detection. Immunoblot analysis revealed that IgG antibodies are mainly reactive with p 100 and the internal flagellin fragment (sensitivity 51% and 32%, respectively) and rarely with OspC (14%). All patients with late Lyme borreliosis had IgG antibodies against the p100. IgM antibodies were predominantly directed against OspC (43%) and in a lower extent against the internal flagellin fragment and p100 (15% and 13%, respectively). The complete flagellin was not useful due to a high number of unspecific reactions with control sera and the OspA was only exceptionally reactive in Lyme borreliosis patients. The sensitivity of IgM antibody detection could be increased in cases with early Lyme borreliosis from 46% to 65% when the OspC blot was performed in addition to the flagellin ELISA, or from 56% to 65% when performed in addition to the OGP-ELISA. The recombinant blot is, therefore, a valuable diagnostic test to increase sensitivity of early antibody detection and is regarded as a valuable confirmatory test also in late disease.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The ospC gene coding for the outer surface protein OspC and the fla gene coding for the flagellin have been investigated in three different Borrelia burgdorferi sensu lato strains. These strains (the North American strain B31 and the European strains PKo and PBi) derive from various biological sources (lxodes dammini, human skin and human CSF) and belong to three different B. burgdorferi OspA serotypes and genospecies (OspA serotype 1, B. burgdorferi sensu stricto; OspA serotype 2, group VS461 and OspA serotype 4, B. garinii, respectively). The ospC and fla genes of the respective strains have been amplified by polymerase chain reaction, cloned in pUC8 and sequenced. The fla as well as the ospC genes were different among the three strains investigated. In general the fla genes are more conserved than the ospC genes. The fla genes have the same length of 1008 nucleotides coding for proteins of 336 amino acids, whereas the ospC genes differ in length. The ospC genes of strains B31, PKo and PBi have 630, 636 and 621 nucleotides encoding proteins of 210, 212 and 207 amino acids, respecctively. The ospC genes exhibit sequence identities between 70% and 74% among each other, sequence identities of the fla genes are in the range 96–97%. The ospC genes could be expressed in Escherichia coli to obtain proteins with and without leader peptides. The expression of the fla gene and an internal gene fragment resulted in the complete flagellin protein and a truncated protein (amino acids 129–251). The different ospC and fla gene products were immunoreactive with monoclonal antibodies and human sera and, thus, enlarge the spectrum of recombinant antigens to improve antibody detection in patients with Lyme borreliosis.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1831
    Keywords: Key words Lyme borreliosis ; Lyme arthritis ; Polymerase chain reaction ; Outer surface protein A ; Borrelia burgdorferi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Borrelia burgdorferi sensu lato, the etiological agent of Lyme borreliosis, has been divided into three genospecies: B. burgdorferi sensu stricto (OspA-type 1), B. afzelii (OspA-type 2) and B. garinii (OspA-type 3–7). Whereas in Europe B. afzelii (OspA-type 2) is predominant among human skin isolates and B. garinii (OspA-type 3–7) among human CSF isolates, some previous serological studies suggested that Lyme arthritis is also associated with B. burgdorferi sensu stricto in Europe. In the present study we designed ospA type-specific PCRs and identified four different ospA types associated with Lyme arthritis. Our study group consisted of 20 patients with positive serology (ELISA and immunoblotting) and clinical criteria for Lyme arthritis. B. burgdorferi DNA was detected in 13 patients and in none of 10 control patients from synovial fluid. We identified ospA-type 1 (26.6%), ospA-type 2 (33.3%), ospA-type 4 (6.6%) and ospA-type 5 (33.3%). Our conclusion is that in Europe B. burgdorferi sensu lato strains causing Lyme arthritis are considerably heterogeneous and that there is no prevalence of certain genospecies or OspA-types among this strains.
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  • 5
    ISSN: 1432-1831
    Keywords: Borrelia burgdorferi, afzelii, garinii ; Outer surface protein A ; ospA genes ; ospA sequence analysis ; Outer surface protein A serotypes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The genes coding for the outer surface protein A (OspA) of 19 different Borrelia burgdorferi strains belonging to the seven OspA-serotypes 1–7, previously described [Wilske et al. (1993) J Clin Microbiol, 31: 340–350], have been investigated. B. burgdorferi sensu lato strains were chosen from various biological sources (ticks, human skin and cerebrospinal fluid) as well as different geographical origins (Germany, Slovenia, Austria, United States). The open reading frames of all ospA genes consist of 819–825 nucleotides corresponding to proteins of approximately 30 kDa. The ospA sequences obtained in this study and previous published studies were compared with the results from OspA serotyping with monoclonal antibodies. The classification into the seven OspA serotypes could be confirmed on a genetic basis (ospA genotypes 1–7) for all strains analyzed so far (n=29). In addition, one strain without OspA expression could be assigned to ospA genotype 2. Genetic stability could be proven for the ospA gene of B. burgdorferi strain PWudI after inocculation and reisolation from a gerbil. However, we found evidence for intragenic recombination by cluster analysis of ospA sequence data. Accordance of ospA genotype 1 strains with B. burgdorferi sensu stricto and ospA genotype 2 strains with B. afzelii, as well as the ospA genotype strains 3–7 with B. garinii was confirmed by pulsed-field gel electrophoresis of MluI-digested genomic DNA. B. garinii is not only more heterogenous in respect to the OspA-encoding genes, but shows moreover major subgroups formed by genotypes 4, 5 and 6 and genotypes 3 and 7, respectively. The latter group has not been described previously and is specifically recognized by an OspA-specific monoclonal antibody L32 1F7.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1831
    Keywords: Key words  Lyme borreliosis  ;  Borrelia burgdorferi  ; Cerebrospinal fluid  ;  OspA  ;  OspC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract   Neuroborreliosis is the most frequent manifestation of the second stage of Lyme borreliosis in Europe. However, only few isolates from the cerobrospinal fluid (CSF) have been characterized with controversial results. A large panel of 36 CSF isolates isolated over a 10-year period in Munich has now been analyzed for their OspA and OspC type, resulting in at least eight different types, respectively. Representatives of the different types cultivated from CSF in Munich have also been isolated from other geographical regions in Europe from CSF or ticks, suggesting a widespread distribution of pathogenic strains. A certain OspA type (type 4) was frequently observed in adults but rarely in children or ticks. Since OspA and OspC are the most promising candidates for a Borrelia vaccine, the considerable heterogeneity found among CSF isolates has important implications for development of a vaccine in Europe.
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  • 7
    ISSN: 1432-1831
    Keywords: Key words  Borrelia burgdorferi sensu lato ; Lyme borreliosis ; Recombinant antigens p100 ; p41/intern ; Enzyme-linked immunosorbent assays ; Antibody prevalence in forest workers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract   To improve IgG antibody detection in the serodiagnosis of Borrelia burgdorferi infection, indirect enzyme-linked immunosorbent assays (ELISAs) were developed utilizing purified recombinant antigens of B. burgdorferi sensu lato: the chromosomally encoded proteins p100 of strain PKo (B. afzelii) and p4li (internal flagellin fragments) derived from strains PKo, PBi (B. garinii), and B31 (B. burgdorferi sensu stricto). In Western blot analysis, these proteins have proved to be highly specific and sensitive for IgG antibody detection, especially in late manifestations of Lyme borreliosis. Sera from 464 forest workers, a high-risk group for infections with B. burgdorferi, were investigated and the results compared with those obtained by a commercial ELISA using an octyl-β-d-glucopyranoside (OGP) extract from B. afzelii (PKo) cells as the antigenic substrate. Sera derived from 200 blood donors served for determination of the 95% specific cut-off level. The number of positive test results using OGP-ELISA (23.9%) was only slightly higher than that using p100-ELISA (19.8%); corresponding results were observed in 84.7% of all sera (14.2% positive, 70.5% negative). The frequency and interassay correspondence of positive results increased with the age of the forest workers, as most markedly demonstrated by p100 and OGP assays (P〈0.0001). Using the p41i-ELISAs, generally no significant strain-dependent differences in sensitivity were found (PKo, 13.8%; PBi, 14.2%; B31, 12.9%). Compared with the p100-ELISA, the p41i-assays showed significantly lower detection rates for IgG antibodies (P〈0.03) and a reduced capacity for quantitative discrimination between seropositive individuals and negative controls (P〈0.0007). At a 95% specificity, the IgG antibody detection rate could be increased to 23.1% when the p100-ELISA was evaluated in combination with the assays using p41i/PBi and P41i/B31. Due to its high sensitivity, the specific recombinant p100-ELISA might be a suitable test for detection of late immune responses to B. burgdorferi, thus being especially useful for epidemiological investigations.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-1831
    Keywords: Key words Human granulocytic ehrlichiosis ; Granulocytic ehrlichiae ; Ixodes ricinus ; Borrelia burgdorferi ; Southern Germany
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Human granulocytic ehrlichiosis (HGE) is an emerging infectious disease recognized in the Western hemisphere. HGE is well known to occur in North America, but records from outside the United States are sparse. The great majority of data from Europe are restricted to seroprevalence studies and molecular biological detection of granulocytic ehrlichiae (GE) in ticks and mammals, but include defined cases from Slovenia. They argue for the existence of this disease in many parts of Europe. In the present study, 510 Ixodes ricinus ticks collected in five different regions of Southern Germany were investigated for the presence of GE and Borrelia burgdorferi sensu lato using polymerase chain reaction. In all, 8 (1.6%) of the 492 ticks that could be evaluated (193 females, 208 males, and 91 nymphs) contained GE and 178 (36.2%) B. burgdorferi s.l.. Four of these ticks were infected with both pathogens. Interestingly, all ehrlichia-infected ticks were adults and all were collected in the English Garden, a recreational park area located in the city of Munich. Sequencing of the 16S rDNA (bp 1–1101) of four of the GE showed 100% sequence identity to each other and greater than 99.9% identity with the published sequence of the HGE agent. The four GE differed in respect to other hitherto described GE by a nucleotide exchange at position 336. These results show that GE that are closely related to the HGE agent are present in Southern Germany, and that coinfection with B. burgdorferi is common in GE-infected ticks. However, in contrast to B. burgdorferi which is endemic everywhere in Southern Germany, the distribution of GE seems to be focal.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-1831
    Keywords: Key words Outer surface protein ; Osp17 ; Borrelia burgdorferi ; Borrelia afzelii ; Lyme borreliosis ; Serodiagnosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Western blot analyses of the human humoral response of patients with Lyme borreliosis have shown that a 17-kDa protein is an immunodominant protein in late disease. Immune electron microscopy with a monoclonal antibody against this protein revealed that the 17-kDa protein is abundantly expressed on the surface of Borrelia afzelii strain PKo. Therefore, the protein has been renamed outer surface protein (Osp)17. Recombinant Osp17 of strain PKo was expressed in Escherichia coli and purified by chromatography. Immunoblot analysis of human sera showed a comparable sensitivity with recombinant and natural proteins. The DNA sequences of the osp17 genes from different B. afzelii strains were determined. The DNA sequences of the different osp17 homologues (six isolates from skin, three isolates from CSF and one isolate from synovial fluid) had high sequence identities of at least 94%. Using a polyclonal antibody against recombinant Osp17, it was shown that Osp17 expression varied considerably among the investigated B. afzelii strains. As previously also observed for OspA- and OspC-encoding genes, the osp17 gene is present in strains not expressing the respective protein. It has been shown that OspA and OspC expression varies in different environments such as tick and vertebrate host. Studies are underway to examine whether this is also true for Osp17. For diagnostic purposes the use of recombinant Osp17 has the advantage that the amount of Osp17 antigen can be easily standardized for immunoblotting, and that this antigen can be used in a protein-specific enzyme-linked immunosorbent assay.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    European journal of pediatrics 157 (1998), S. 953-954 
    ISSN: 1432-1076
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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