Evaluation of the syphilis Bio-EnzaBead assay for detection of treponemal antibody.

An enzyme-linked immunosorbent assay for Treponema pallidum antibody (Syphilis Bio-EnzaBead Kit; Organon Teknika Corp., formerly Litton Bionetics, Kensington, Md.) was compared with the standard fluorescent treponemal antibody absorption test for syphilis (indirect fluorescent-antibody confirmatory test; Zeus Scientific Inc., Raritan, N.J.). Six hundred specimens tested in the rapid plasma Reagin card test (Hynson, Westcott and Dunning, Inc., Baltimore, Md.) and microhemagglutination assay for antibodies to T. pallidum (Ames Division, Miles Laboratories, Inc., Elkhart, Ind.) were used for comparison testing. One hundred and sixty-two specimens were from either persons with syphilis or persons whose history was unknown but who showed reactive serology, whereas 438 were from persons without syphilis. The reactivity of each serum by the Bio-EnzaBead test was determined visually and from optical density readings. Excellent reproducibility of visual readings was obtained; all results were within +/- 1 gradation of the expected readings in all 60 coded sera tested. Overall agreement between the Bio-EnzaBead and fluorescent treponemal antibody absorption test results was 96.3% when read visually and greater than or equal to 95.7% when using optical density readings. Our data indicate that Bio-EnzaBead results read at 405 nm and determined by using the mean antigen optical density reading of the nonreactive control plus 0.025 as the cutoff value provide an overall sensitivity and specificity of 93.0 and 98.6%, respectively, along with the best agreement with the fluorescent treponemal antibody absorption assay. Ease of performance and objectivity also contribute toward the acceptability of this assay as an alternative confirmatory test for syphilis.