Evaluation of a Commercial Multiplex Quantitative PCR (qPCR) Assay for Simultaneous Detection of Mycoplasma genitalium and Macrolide Resistance-Associated Mutations in Clinical Specimens

Macrolide antibiotics are the first-line treatment for Mycoplasma genitalium infections; however, macrolide resistance has increased up to 40% in several countries (1–3). Consequently, the 2016 European guideline on M. genitalium infections has recommended complementing the molecular detection of M. genitalium with an assay capable of detecting macrolide resistance-associated mutations (4). We aimed to evaluate the clinical performance of the CE-marked ResistancePlus MG kit (SpeeDx, Australia), which utilizes PlexZyme/PlexPrime technology (5) for the detection of M. genitalium (MgPa adhesin gene) and the five predominant 23S rRNA macrolide resistance-associated mutations (A2058G, A2059G, A2058C, A2059C, and A2058T [Escherichia coli numbering]). This was compared to in-house assays using realtime quantitative PCR (qPCR) to detect M. genitalium and real-time PCR and melting curve analysis to detect the macrolide resistance-associated mutations (“macrolide resistance qPCR”) (6). A total of 206 male and female urogenital specimens previously analyzed using an in-house method (7) and conserved at 80°C (94 M. genitalium-positive and 112 M. genitalium-negative specimens) were retrospectively and systematically selected from samples collected in 2014 to 2015 at the Bordeaux University Hospital (France). A 5l volume of the internal control provided in the kit was spiked into 200 l of specimen before extraction, which was performed using a MagNA Pure 96 instrument (Roche Diagnostics). The ResistancePlus MG and in-house assays were performed using the Cobas z480 analyzer of the Cobas 4800 platform (Roche Diagnostics), according to the instructions of the manufacturers. An additional M. genitalium detection assay, using the CE-marked S-DiaMGTV kit (Diagenode, Belgium) (8, 9), was performed such that any two of the possible three comparator results would define infection status. PyroseAccepted manuscript posted online 28 December 2016 Citation Le Roy C, Hénin N, Bébéar C, Pereyre S. 2017. Evaluation of a commercial multiplex quantitative PCR (qPCR) assay for simultaneous detection ofMycoplasma genitalium and macrolide resistance-associated mutations in clinical specimens. J Clin Microbiol 55:978–979. https://doi.org/10.1128/ JCM.02168-16. Editor Robin Patel, Mayo Clinic Copyright © 2017 Le Roy et al. This is an openaccess article distributed under the terms of the Creative Commons Attribution 4.0 International license. Address correspondence to Sabine Pereyre, [email protected].