Lyme Disease: Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection and Quantitation of Antibodies Against Borrelia burgdorferi

Abstract

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantitation of Borrelia burgdorferi antibodies. When compared with IFA and FIAX (Quantitative Fluorescence Immunoassay), this newly developed ELISA demonstrated equivalent sensitivity (97.5%) and specificity (95.1%). Linearity of the ELISA was established using serial two-fold dilutions of serum, thus allowing the use of a single dilution of serum for quantitation of antibody. Positive values were reliable with a mean intra-assay coefficient of variation of 4.30% and a mean inter-assay coefficient of variation of 9.44%. The antigen coated solid phase was shown to be stable for at least three months when stored at 4°C. Sera containing Rheumatoid Factor, anti-nuclear antibodies and antibodies against Rocky Mountain spotted fever did not react in this assay. One of several sera reactive with Treponema pallidum, also a spirochete, exhibited minimal reactivity in the Borrelia burgdorferi ELISA. In overall performance, the ELISA demonstrated the sensitivity, specificity and accuracy essential for use as a diagnostic tool for the detection and quantitation of antibodies to Borrelia burgdorferi.