Use of a hybrid protein consisting of the variable region of the Borrelia burgdorferi flagellin and part of the 83-kDa protein as antigen for serodiagnosis of Lyme disease.
A hybrid protein consisting of the variable region of the Borrelia burgdorferi flagellin (an 18-kDa fragment) and a 59-kDa fragment (lacking the N-terminal part) of the 83-kDa protein has been constructed by genetic engineering. It was expressed as a nonfusion protein of an apparent molecular weight of 77,000 in Escherichia coli. The suitability of this new antigen for the diagnosis of Lyme disease was tested by immunoblotting; for comparison, the recombinant variable region of the flagellin, the 18-kDa fragment (p18), and the whole recombinant 83-kDa protein (p83), both expressed in E. coli, were used. A total of 120 serum samples from various stages of Lyme disease, which were positive in two serological assays, a passive hemagglutination assay and an indirect immunofluorescence assay, were tested. By indirect immunofluorescence, 74 samples were positive for immunoglobulin G (IgG) antibodies and 72 were positive for IgM antibodies. Of these serum samples, 69 of 74 (93%) contained IgG antibodies against p18 and/or p83, and IgG antibodies were detected by the hybrid protein in 67 (90%) samples. IgM antibodies against p18 and/or p83 were detected in 60 of 72 (83%) serum samples, and 57 (79%) serum samples were reactive with the hybrid protein. Twenty serum samples of patients with a history of syphilis and 40 serum samples, negative in routine B. burgdorferi serology, were tested as controls. The hybrid protein, made up of specific epitopes of an early (p18) and late (p83) antigen, is recognized by almost the same number of patient serum samples as the individual antigens.
Institut für Medizinische Mikrobiologie und Hygiene, Freiburg, Germany.
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C
Rasiah
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Rauer
G S
GS
Gassmann
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A
Vogt
D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005408:Flagellin / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D005798:Genes, Bacterial; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics; D008193:Lyme Disease / Q000175:diagnosis* / Q000276:immunology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012698:Serologic Tests
D000942:Antigens, Bacterial; D017931:DNA Primers; D016056:Immunodominant Epitopes; D011994:Recombinant Proteins; D005408:Flagellin