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  • 1
    Electronic Resource
    Electronic Resource
    Recombinant immunoblot in the serodiagnosis of Lyme borreliosis (1993)
    Springer
    Medical microbiology and immunology 182 (1993), S. 255-270 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00579624
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  • 2
    Electronic Resource
    Electronic Resource
    Expression of outer surface proteins A and C of Borrelia burgdorferi in Ixodes ricinus ticks removed from humans (1998)
    Springer
    Medical microbiology and immunology 187 (1998), S. 121-126 
    Details
    ISSN: 1432-1831
    Keywords: Key words Outer surface protein expression ; Borrelia burgdorferi ; Ticks (Ixodes ricinus) ; Lyme disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A total of 131 Ixodes ricinus (51 females, 1 male and 79 nymphs) removed from persons living in Southern Germany were investigated by immunofluorescence assay for the presence of Borrelia burgdorferi with a polyvalent rabbit immune serum and monoclonal antibodies specific for outer surface proteins (Osp) A or C. Borreliae were detectable in 48 (36.6%) of the ticks. Infection rates of these adults and nymphs were significantly higher than infection rates of unfed ticks from Southern Germany. Borreliae in 31.3% (n = 15) of the infected ticks expressed solely OspA, solely OspC in 12.5% (n = 6), and both OspA and OspC in 39.6% (n = 19) of ticks, while in 16.7% (n = 8) of ticks neither were expressed. Presentation of OspC by B. burgdorferi in I. ricinus was correlated with tick weight: in females, OspC was detectable only in ticks with a minimum weight of about 3.5 mg, and in nymphs weighing at least 1 mg. These results indicate that in I. ricinus removed from humans OspC is up-regulated during the blood meal of the tick, but in most ticks OspA is still detectable and might even be present in the absence of OspC expression in the midgut and salivary glands of nearly fully engorged nymphal ticks. Furthermore, we found strong evidence that borreliae expressing solely OspA while in the salivary glands can cause Lyme borreliosis. Our findings indicate that during tick feeding, humans are exposed to borreliae that may express either OspA or OspC or both, or lack both OspA and C. These findings suggests that, at the minimum, both OspA and C should be considered as vaccine candidates for prophylaxis of Lyme borreliosis in Europe.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004300050083
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  • 3
    Electronic Resource
    Electronic Resource
    Immunoblot using recombinant antigens derived from different genospecies of Borrelia burgdorferi sensu lato (1994)
    Springer
    Medical microbiology and immunology 183 (1994), S. 43-59 
    Details
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunodominant proteins are variable in molecular and antigenic structure among different genospecies of Borrelia burgdorferi sensu lato. We have recently developed an immunoblot using five recombinant antigens: the chromosomal-encoded B. burgdorferi proteins p 100, the flagellin and an internal flagellin fragment thereof, and the plasmid-encoded outersurface proteins A (OspA) and C (OspC). In the present study the same antigens (derived from strain PKo, genospecies B. afzelii) were compared with the homologous recombinant proteins from strain B31 (genospecies B. burgdorferi sensu stricto) and with OspA, OspC and the internal flagellin fragment from strain PBi (genospecies B. garinii). Patients with neuroborreliosis (n=28) and patients with acrodermatitits chronica atrophicans (n=20) were investigated in the IgG immunoblot; the IgM immunoblot was performed only in patients with neuroborreliosis. There was a small increase in the detection rate of OspA-specific IgG or IgM antibodies using the different variants of recombinant OspA; however, OspA remained an insensitive antigen for antibody detection in Lyme borreliosis. The same was true to OspC-specific IgG antibodies. The sensitivity of OspC, which is the immunodominant antigen for IgM antibody detection, could not be increased using recombinant antigens derived from different strains. However, some sera which were negative in the recombinant immunoblot reacted with OspC in the conventional immunoblot using B. burgdorferi whole cell lysate as antigen. The most unexpected finding was the high degree of immunological heterogeneity of the internal flagellin fragments: IgG antibodies were detected in 18 of 48 patients using B31 fragments, in 25 of 48 using PKo fragments, in 23 of 48 using PBi fragments versus 33 of 48 when the three recombinant proteins were combined. PKo-derived fragments were more sensitive for antibody detection in patients with acrodermatitis chronica atrophicans, B31- and PBi-derived fragments for antibody detection in patients with neuroborreliosis. This is in agreement with the fact that isolates from patients with neuroborreliosis are predominantly belonging to the genospecies B. burgdorferi sensu stricto and B. garinii. For detection of IgM antibodies in sera from patients with neuroborreliosis, recombinant internal fragments derived from strains B31 and PBi were more sensitive than the PKo-derived fragment. The best discrimination between neuroborreliosis sera and control sera was achieved when the IgM blot was performed using recombinant internal flagellin fragments derived from strains PKo and PBi and OspC derived from B31 or PKo.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00193630
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  • 4
    Electronic Resource
    Electronic Resource
    Diversity of OspA and OspC among cerebrospinal fluid isolates of Borrelia burgdorferi sensu lato from patients with neuroborreliosis in Germany (1996)
    Springer
    Medical microbiology and immunology 184 (1996), S. 195-201 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004300050007
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  • 5
    Electronic Resource
    Electronic Resource
    Coexistence of ehrlichiae of the phagocytophila group with Borrelia burgdorferi in Ixodes ricinus from Southern Germany (1999)
    Springer
    Medical microbiology and immunology 188 (1999), S. 145-149 
    Details
    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
    URL: http://dx.doi.org/10.1007/s004300050117
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  • 6
    Electronic Resource
    Electronic Resource
    Immunological and molecular variability of OspA and OspC. implications forBorrelia vaccine development (1996)
    Springer
    Infection 24 (1996), S. 208-212 
    Details
    ISSN: 1439-0973
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Borrelia burgdorferi sensu lato, der Erreger der Lyme-Borreliose, ist in Europa ausgesprochen heterogen. Da die äußeren Membranproteine OspA und OspC die Hauptkandidaten für eine Borrelien-Vakzine sind, wurde deren immunologische und molekulare Heterogenität untersucht. Aufgrund von Westernblotanalysen mit monoklonalen Antikörpern und Sequenzanalysen von PCR-amplifiziertem OspA und OspC fanden wir sieben verschiedene OspA- und 16 verschiedene OspC-Typen. Während Hautisolate (n=68) ganz vorwiegend (84%) OspA-Serotyp 2 (oderBorrelia afzelii) angehörten, waren Isolate aus Liquor cerebrospinalis und von Zecken (n=43 bzw. n=90) ausgesprochen heterogen im OspA-Typ mit Prävalenz der mitBorrelia garinii assoziierten OspA-Typen. OspA-Typ 4 fand sich oft bei Liquorisolaten (28%). Von Zecken wurde OspA-Typ 4 bisher nicht isoliert. Wie in einer anderen Studie berichtet, konnte jedoch Typ 4-OspA in Zecken mit der hochsensitiven PCR nachgewiesen werden.
    Notes: Summary Borrelia burgdorferi sensu lato, the etiological agent of Lyme borreliosis is considerably heterogeneous in Europe. Since the outer surface proteins OspA and OspC are the most promising candidates for aBorrelia vaccine the immunological heterogeneity of these proteins was investigated. By immunological analysis with monoclonal antibodies and sequence analysis of PCR amplified OspA and OspC at least seven and 16 different types, respectively, were found. Whereas skin isolates (n=68) were quite homogeneous (84% belonged to OspA-serotype 2 orBorrelia afzelii), isolates from human cerebrospinal fluid and from ticks (n=43 and n=90 respectively) were highly heterogeneous in their OspA-serotypes with prevalence of theBorrelia garinii associated types (about 70%). OspA-type 4 was often found among isolates from cerebrospinal fluid (28%). In ticks type 4 OspA has not been detected by culture so far. However, as reported in a previous study, type 4 OspA could be detected in ticks by the highly sensitive PCR technique.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01713341
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