Enzyme-linked immunosorbent assay for detection of specific antibodies to Ureaplasma urealyticum serotypes

Abstract

Optimal conditions of a micro-enzyme-linked immunosorbent assay system for the detection of immunoglobulin G antibodies to Ureaplasma urealyticum were established with rabbit antisera. Initially, the antisera, raised against eight U. urealyticum serotypes grown on medium containing horse serum, displayed nonspecific reactions with our enzyme-linked immunosorbent assay antigens. Substitution of fetal bovine serum in the medium eliminated this nonspecificity. The assay was then serotype-specific for the original eight U. urealyticum serotypes. The prominent homologous reaction was easily differentiated from the heterologous reactions. A one-way cross-reaction was observed with serotype 2 antiserum and serotype 5 antigen. The results were reproducible and could be obtained in 4 h with only 10 microliters of serum for eight serotypes. Optimal antigen concentrations for the U. urealyticum serotypes ranged from 0.40 to 1.60 micrograms/ml. Our results indicated that enzyme-linked immunosorbent assay has the potential for the detection of antibodies to specific serotypes of U. urealyticum.