Abstract
We have found that sera from patients with early stages of Lyme disease contain predominant immunoglobulin M reactivity to a major 23-kDa protein (p23) from Borrelia burgdorferi 2591 isolated in Connecticut. To characterize this immunodominant antigen, we cloned and sequenced p23 and found it to be 83% identical by nucleotide sequence and 75% identical by amino acid sequenced to pC (recently renamed OspC), an abundantly expressed protein on the outer surface of PKo, a European strain of B. burgdorferi (B. Wilske, V. Preac-Mursic, S. Jauris, A. Hofmann, I. Pradel, E. Soutschek, E.Schwab, G. Will, and G. Wanner, Infect. Immun. 61:2182-2191, 1993). In addition, immunoelectron microscopy localized p23 to the outer membrane, confirming that p23 is the strain 2591 homolog of OspC. The North American strain B31, commonly used in serologic assays for Lyme disease, does not express OspC. Northern (RNA) blot analysis detected low levels of ospC mRNA in B31, and DNA sequencing of the ospC gene from B31 revealed a 54-bp deletion in the upstream regulatory region, possibly accounting for the low transcriptional activity of ospC. The ospC coding region from B31 was cloned and antibody-reactive OspC was expressed in Escherichia coli. An immunoglobulin M enzyme-linked immunosorbent assay using recombinant OspC as the target antigen shows promise for the serodiagnosis of early stages of Lyme disease.
MeSH Terms
Amino Acid Sequence
Antibodies, Bacterial/biosynthesis
Antibodies, Monoclonal
Antigens, Bacterial
Bacterial Outer Membrane Proteins
Bacterial Proteins/genetics,immunology
Base Sequence
Borrelia burgdorferi Group/genetics,immunology,isolation & purification
Cloning, Molecular
DNA, Bacterial/genetics
Enzyme-Linked Immunosorbent Assay/methods
Gene Expression
Genes, Bacterial
Humans
Immunoglobulin M/biosynthesis
Lyme Disease/diagnosis,immunology,microbiology
Microscopy, Immunoelectron
Molecular Sequence Data
North America
Recombinant Fusion Proteins/genetics,immunology
Sequence Homology, Amino Acid
Transcription, Genetic
Chemicals
Antibodies, Bacterial
Antibodies, Monoclonal
Antigens, Bacterial
Bacterial Outer Membrane Proteins
Bacterial Proteins
DNA, Bacterial
Immunoglobulin M
OspC protein
Recombinant Fusion Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Padula S J
Department of Medicine, University of Connecticut Health Center, Farmington 06030-1310.
Sampieri A
Dias F
Szczepanski A
Ryan R W
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