Abstract
Background
Performance of a 16S rRNA analysis of the cervicovaginal microbiome of 220 participants recruited into the T Cell Response against Chlamydia (TRAC) cohort between February 2011 and August 2014 in Allegheny County, Pennsylvania USA surprisingly detected DNA encoding chlamydial 16S rRNA in samples from seven participants who had tested negatively for Chlamydia trachomatis (CT) and DNA encoding gonococcal 16S rRNA from five participants who had tested negatively for Neisseria gonorrhoeae (NG) infection with the Aptima Combo2 assay (Hologic).
Methods
We used targeted PCR amplification followed by sequencing to characterize the chlamydial 23S rRNA locus and qPCR to detect gonococcal DNA in residual diagnostic swab eluates or DNA used to generate 16S rRNA libraries.
Results
Discrepant specimens that contained chlamydial DNA carried a diagnostic-avoidant, G1526A variant in the 23S rRNA locus identical to variants previously detected in Finland, Denmark, and the UK. PCR validation of gonococcal DNA was confirmed for all participants who had tested negatively, with stochastic effects consistent with infection levels close to the limit of detection by the diagnostic assay.
Conclusions
These data indicate that this probe-avoidant CT mutant, and possibly others, were circulating in the northeastern US prior to their detection and characterization in 2019 and subsequently. Although infrequent, documentation of false negative results for CT indicates a need for clinicians to consider performance of a second test that uses alternate PCR probes if patients have persistent symptoms or have known contact to an infected sex partner and their initial NAAT is negative.
Full Text Availability
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