Cloning of the gene encoding the decorin-binding protein B (DbpB) in Borrelia burgdorferi sensu lato and characterisation of the antibody responses to DbpB in Lyme borreliosis.
A genome walking technique was applied to borrelial DNA to clone the gene encoding decorin-binding protein B (DbpB) in Borrelia garinii and B. afzelii. Sequence analysis showed 62-67% identity of the predicted amino acid sequences of DbpB between the B. afzelii and B. garinii strains and B. burgdorferi sensu stricto. Within subspecies, the sequences were 99-100% identical. The respective recombinant DbpBs (rDbpBs) were produced and tested as antigens in an enzyme-linked immunosorbent assay (ELISA) for Lyme borreliosis (LB). In IgG ELISA, with rDbpBs as antigens, 11 (73%) of 15 adult patients with Lyme arthritis and 9 (64%) of 14 with neuroborreliosis were positive. Of children with Lyme arthritis, 40 (77%) of 52 were positive. All adult and paediatric patients with disseminated LB had high titres of anti-flagellin IgG antibodies. Seropositivity against rDbpB from B. garinii predominated, 39 (65%) of 60 of the positive samples reacting with rDbpB from B. garinii. In patients with erythema migrans, IgM antibodies to rDbpB were detected in 1 (4%) of 23 and IgG antibodies in 6 (26%) of 23. These results indicate that DbpB may be a useful antigen in the IgG serology for disseminated LB. The high inter-species sequence heterogeneity observed indicates that a combination of the variant DbpBs should be included in the antigen set to cover all the relevant borrelial subspecies in the serodiagnosis of LB.
Heikkilä
Tero
T
*Hospital for Children and Adolescents, University of Helsinki, Stenbäckinkatu 11, FIN-00290 Helsinki, †Department of Bacteriology and Immunology, Haartman Institute, Haartmaninkatu 3, FIN-00290 Helsinki, ‡Laboratory Diagnostics, Helsinki University Central Hospital, Haartmaninkatu 3, FIN-00290 Helsinki, §Department of Otorhinolaryngology, University of Helsinki, 00290 Helsinki, Finland and Professor- Hess-Children's Hospital, Bremen Central Hospital, 28205 Bremen, Germany.
Seppälä
Ilkka
I
*Hospital for Children and Adolescents, University of Helsinki, Stenbäckinkatu 11, FIN-00290 Helsinki, †Department of Bacteriology and Immunology, Haartman Institute, Haartmaninkatu 3, FIN-00290 Helsinki, ‡Laboratory Diagnostics, Helsinki University Central Hospital, Haartmaninkatu 3, FIN-00290 Helsinki, §Department of Otorhinolaryngology, University of Helsinki, 00290 Helsinki, Finland and Professor- Hess-Children's Hospital, Bremen Central Hospital, 28205 Bremen, Germany.
Saxen
Harri
H
*Hospital for Children and Adolescents, University of Helsinki, Stenbäckinkatu 11, FIN-00290 Helsinki, †Department of Bacteriology and Immunology, Haartman Institute, Haartmaninkatu 3, FIN-00290 Helsinki, ‡Laboratory Diagnostics, Helsinki University Central Hospital, Haartmaninkatu 3, FIN-00290 Helsinki, §Department of Otorhinolaryngology, University of Helsinki, 00290 Helsinki, Finland and Professor- Hess-Children's Hospital, Bremen Central Hospital, 28205 Bremen, Germany.
Panelius
Jaana
J
*Hospital for Children and Adolescents, University of Helsinki, Stenbäckinkatu 11, FIN-00290 Helsinki, †Department of Bacteriology and Immunology, Haartman Institute, Haartmaninkatu 3, FIN-00290 Helsinki, ‡Laboratory Diagnostics, Helsinki University Central Hospital, Haartmaninkatu 3, FIN-00290 Helsinki, §Department of Otorhinolaryngology, University of Helsinki, 00290 Helsinki, Finland and Professor- Hess-Children's Hospital, Bremen Central Hospital, 28205 Bremen, Germany.
Peltomaa
Miikka
M
*Hospital for Children and Adolescents, University of Helsinki, Stenbäckinkatu 11, FIN-00290 Helsinki, †Department of Bacteriology and Immunology, Haartman Institute, Haartmaninkatu 3, FIN-00290 Helsinki, ‡Laboratory Diagnostics, Helsinki University Central Hospital, Haartmaninkatu 3, FIN-00290 Helsinki, §Department of Otorhinolaryngology, University of Helsinki, 00290 Helsinki, Finland and Professor- Hess-Children's Hospital, Bremen Central Hospital, 28205 Bremen, Germany.
Huppertz
Hans-Iko
HI
Lahdenne
Pekka
P
*Hospital for Children and Adolescents, University of Helsinki, Stenbäckinkatu 11, FIN-00290 Helsinki, †Department of Bacteriology and Immunology, Haartman Institute, Haartmaninkatu 3, FIN-00290 Helsinki, ‡Laboratory Diagnostics, Helsinki University Central Hospital, Haartmaninkatu 3, FIN-00290 Helsinki, §Department of Otorhinolaryngology, University of Helsinki, 00290 Helsinki, Finland and Professor- Hess-Children's Hospital, Bremen Central Hospital, 28205 Bremen, Germany.
D018829:Adhesins, Bacterial; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis* / Q000097:blood; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins; D001483:Base Sequence; D015748:Borrelia burgdorferi Group / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology; D002648:Child; D016386:Chromosome Walking; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis; D008193:Lyme Disease / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; C112557:DdpB protein, Borrelia burgdorferi; D007074:Immunoglobulin G; D011994:Recombinant Proteins