Evaluation of Gen-Probe APTIMA-based Neisseria gonorrhoeae and Chlamydia trachomatis confirmatory testing in a metropolitan setting of high disease prevalence

Prompted by reports challenging the validity of the low-positive Neisseria gonorrhoeae and Chlamydia trachomatis results generated by the APTIMA Combo 2 assay (Gen-Probe, Incorporated) and by a Centers for Disease Control and Prevention recommendation to confirm N. gonorrhoeae- or C. trachomatis-positive screens by using an alternative amplification target, we report on a comparison of this means of confirmation with an in-house algorithm of repeat testing. Primary clinical specimens yielding N. gonorrhoeaeor C. trachomatis-specific luminescent values between 100,000 and 1,000,000 were repeat tested in duplicate. A subset of specimens was forwarded for confirmatory assays (Gen-Probe) individualized for alternative N. gonorrhoeae or C. trachomatis targets. An 18-month audit revealed that 230 of 29,977 C. trachomatis screens (0.8%) and 41 of 29,064 N. gonorrhoeae assays (0.1%) yielded low-positive data. When a subset of 40 low-positive N. gonorrhoeae screens was repeat tested, 20 (50.0%) remained positive; 22 (55.0%) of the screens remained positive following performance of the confirmatory assay. In contrast, repeat testing of 153 low-positive C. trachomatis screens yielded a positive result for fewer specimens (n = 97; 63.4%) than when commercial confirmatory testing was used (n = 124; 81.0%). However, confirmation of the results for additional C. trachomatis screens by use of an alternative target did not translate into significant differences in the calculated overall C. trachomatis-positive screen rates (7.39% by repeat testing versus 7.52% by the confirmatory assay; P = 0.53). Furthermore, use of the confirmatory assay raised the positive predictive value only 1.8% over that of repeat testing. Molecular confirmatory testing did not significantly enhance the reliability of C. trachomatis- or N. gonorrhoeae-specific nucleic acid amplification testing in this metropolitan setting compared to the reliability of repeat testing.