- BACKGROUND AND GOAL: The VIDAS Enzyme Linked Fluorescent Assay is a fully automated assay for the detection of Chlamydia trachomatis in clinical specimens. Because there is an increasing demand for automated assays for large volume laboratories and there is little performance data available, the authors evaluated the performance of the VIDAS enzyme linked fluorescent assay by comparing it with Chlamydiazyme and polymerase chain reaction. STUDY DESIGN: Endocervical swabs from 330 women attending a hospital based obstetrics/gynecology clinic and 100 first void urine specimens from males attending a sexually transmitted disease clinic were tested by enzyme linked fluorescent assay, Chlamydiazyme, and polymerase chain reaction. RESULTS: Fourteen of 330 endocervical specimens and 14 of 100 first void urine specimens were positive by Chlamydiazyme. Enzyme linked fluorescent assay detected 12 of 14 endocervical and 11 of 14 first void urine specimens. Compared with Chlamydiazyme, enzyme linked fluorescent assay had a sensitivity of 85.7% (12 of 14) for endocervical swabs and 76.9% (11 of 14) for first void urine specimens. Polymerase chain reaction detected an additional five endocervical and two first void urine specimens that had negative results by both enzyme linked fluorescent assay and Chlamydiazyme. All 7 were confirmed positive by polymerase chain reaction using a second primer set. Using an expanded gold standard of blocked Chlamydiazyme and confirmed polymerase chain reaction, enzyme linked fluorescent assay had a sensitivity of 63.2% (12 of 19) for endocervical swabs and 68.8% (11 of 16) for first void urine specimens compared with 73.7% (14 of 19) and 87.5% (14 of 16) for Chlamydiazyme. Polymerase chain reaction had a sensitivity of 100% (19 of 19) and 93.8% (15 of 16) for endocervical swabs and first void urine specimens, respectively. The specificity of enzyme linked fluorescent assay and Chlamydiazyme was 100%. CONCLUSIONS: The VIDAS enzyme linked fluorescent assay for the detection of Chlamydia trachomatis in genitourinary specimens is highly specific but is not sufficiently sensitive for use as a routine diagnostic test.