Report Refuting Value of Immune Complexes To Diagnose Lyme Disease Is Invalid
Abstract
Lyme disease (LD) is difficult to diagnose absent hallmark clinical symptoms. LD immunoassays are problematic because of cross-reactivities, false-positives, sequestration of antibody and antigen in immune complexes (IC) early after infection, and persisting antibodies. Demonstration of IC in LD (10) opened the possibility of developing assays to detect immunoglobulin M (IgM) in IC at a time when antibiotic therapy would be useful. Subsequently, we confirmed the presence or absence of LD antigens/antibodies in sera and other biological fluids from putative LD patients (1-4). Assay development used "gold standard" erythema migrans and/or culture-positive (from biopsy) samples (e.g., CDC) and endemic area controls. We compared our enzyme-linked immunosorbent assay and immunoblots with many types of commercial (e.g., MarDx) and laboratory (e.g., immunofluorescent antibody, enzyme immunoassays) tests. Using monoclonal antibodies (MAb), standard assay development protocols (antibody/antigen checkerboards to ascertain correct concentrations of primary and secondary antibodies), appropriate blocking reagents, and sensitive detection methods (e.g., biotin-avidin), we obtained superior sensitivity and specificity of IC over comparator tests (3, 4) or whole serum alone (corroborated in independent laboratories). IC allowed us to "see" more bands in immunoblots (2) than a comparator commercial test, and exhaustive absorption of IC along with a "good" MAb