New method for detection of Borrelia burgdorferi antigen complexed to antibody in seronegative Lyme disease.
Serologic tests for Lyme disease are problematic. Because of cross-reactive antigens Borrelia burgdorferi (Bb) shares with other organisms, Lyme disease can be overdiagnosed. However, in addition to specificity problems, serologic tests for early Lyme disease can be falsely negative due to lack of sensitivity of ELISAs and Western blots. Most routine antibody tests are designed to detect free antibodies, and in early, active disease, circulating antibodies may not be free in serum but sequestered in complexes with the antigens which originally triggered their production. This difficulty may be overcome by first isolating immune complexes (IC) from the serum and using this fraction for testing. Free Borrelia-specific antibodies can then be liberated from the immune complexes which may enhance test sensitivity in patients with active disease. We developed a technique that captures the antibody component of IC on immunobeads, and subsequently releases the antigen component of IC. Immunoblotting with monoclonal antibody detected at least one antigen to be OspA, thus definitively demonstrating a Borrelia-specific antigen in circulating IC in early Lyme disease. This test is also useful in demonstrating Bb antigen in otherwise seronegative Lyme disease patients.
The Children's Hospital of Philadelphia, Department of Rheumatology, Abramson Research Center 1104D, 3516 Civic Center Blvd., Philadelphia, PA 19104-4318, USA. brunner@email.chop.edu
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Brunner
D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000097:blood / Q000276:immunology*; D015748:Borrelia burgdorferi Group / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D008193:Lyme Disease / Q000097:blood / Q000276:immunology / Q000382:microbiology*; D012680:Sensitivity and Specificity; D012698:Serologic Tests
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial