title: Prospective Evaluation of ResistancePlus MG, a New Multiplex Quantitative PCR Assay for Detection of Mycoplasma genitalium and Macrolide Resistance.
abstract: Mycoplasma genitaliumis a significant pathogen for which first-line treatment is becoming less effective due to increased resistance to macrolides. As conventional culture and antimicrobial susceptibility testing is not feasible for routine detection of this pathogen, molecular markers such as detection of mutations in the 23S rRNA gene have been described to predict resistance. Recently, a novel multiplex quantitative PCR (qPCR) assay, ResistancePlus MG, has been described for the simultaneous detection ofMycoplasma genitaliumand macrolide resistance. In the current study, the clinical performance of the assay was evaluated on 1,089 consecutive urine and anogenital swab samples in symptomatic and asymptomatic male and female patients. Overall, 6.0% were positive forM. genitalium, with 63.1% having macrolide resistance-associated mutations. Compared to the laboratory-validated qPCR method targeting the 16S rRNA gene and Sanger sequencing to determine 23S rRNA mutations, the sensitivity and specificity ofM. genitaliumdetection were 98.5% and 100% and for detection of macrolide resistance mutations were 100.0% and 96.2%, respectively. This assay offers a considerable advantage in clinical settings forM. genitaliumtesting by making the results of macrolide resistance and mutation analyses simultaneously available, which is increasingly important with escalating macrolide resistance.
authors.affiliation: Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia sepehr@unimelb.edu.au.|Department of Obstetrics and Gynaecology, University of Melbourne, Victoria, Australia.|Murdoch Childrens Research Institute, Parkville, Victoria, Australia.
authors.forename: S N
authors.initials: SN
authors.lastname: Tabrizi
authors.affiliation: Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia.|Department of Obstetrics and Gynaecology, University of Melbourne, Victoria, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Su
authors.affiliation: Melbourne Sexual Health Centre, Carlton, Victoria, Australia.|Central Clinical School, Monash University, Frankston, Victoria, Australia.
authors.forename: C S
authors.initials: CS
authors.lastname: Bradshaw
authors.affiliation: Melbourne Sexual Health Centre, Carlton, Victoria, Australia.|Central Clinical School, Monash University, Frankston, Victoria, Australia.
authors.forename: C K
authors.initials: CK
authors.lastname: Fairley
authors.affiliation: SpeeDx Pty Ltd., Sydney, NSW, Australia.
authors.forename: S
authors.initials: S
authors.lastname: Walker
authors.affiliation: SpeeDx Pty Ltd., Sydney, NSW, Australia.
authors.forename: L Y
authors.initials: LY
authors.lastname: Tan
authors.affiliation: SpeeDx Pty Ltd., Sydney, NSW, Australia.
authors.forename: E
authors.initials: E
authors.lastname: Mokany
authors.affiliation: Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia.|Department of Obstetrics and Gynaecology, University of Melbourne, Victoria, Australia.|Murdoch Childrens Research Institute, Parkville, Victoria, Australia.
authors.forename: S M
authors.initials: SM
authors.lastname: Garland
mesh_terms: D001003:Anal Canal / Q000382:microbiology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D024881:Drug Resistance, Bacterial; D005260:Female; D005835:Genitalia / Q000382:microbiology; D006801:Humans; D018942:Macrolides / Q000494:pharmacology; D008297:Male; D008826:Microbial Sensitivity Tests / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009175:Mycoplasma Infections / Q000175:diagnosis* / Q000382:microbiology; D045704:Mycoplasma genitalium / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D011446:Prospective Studies; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014556:Urine / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D018942:Macrolides; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S
keywords: 23S rRNA; Mycoplasma genitalium; NAAT; diagnostic; macrolide resistance; multiplex assay; qPCR; sensitivity and specificity