BACKGROUND: Several nucleic acid amplification (NAA)-based kits are commercially available for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC). A parallel evaluation of the BD ProbeTec ET and the Roche Cobas Amplicor was performed and compared in respect to validity, cost-per-test, hands-on time and turnaround time. METHODS: A total of 703 endocervical swabs were collected at Vanderbilt’s STD clinic. These samples were tested for both CT and GC using the BD Probetec ET, Roche Cobas Amplicor and the GenProbe Pace 2 system. RESULTS: The test results were compared in respect to performance, timeliness, and cost. The positivity rate of the ProbeTec, Cobas Amplicor and Pace 2 was 7.4%, 7.1 % and 5.1% for CT and 1.9%, 1.7% and 1.6% for GC. Using a repeatedly positive result from any three tests as an evaluation standard, the sensitivity of the ProbeTec, Cobas Amplicor and Pace 2 was 96.3%, 92.6% and 66.7% for CT and 100%, 92.3% and 84.6% for GC, respectively. Hands-on time for all three procedures were similar, yet a lower turnaround time (TAT) was noted for the ProbeTec and Pace 2 (4 hours) as compared to the Cobas Amplicor (almost 6 hours). The cost-per test was lower with Pace 2 than either of the amplified tests, Cobas Amplicor and ProbeTec.