Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerations

ABSTRACT Recent work has optimized parameters of a real-time reverse transcriptase PCR-based laboratory-developed test on the Panther Fusion system which detects Mycoplasma genitalium-specific macrolide resistance-associated mutations LDT (MRM-LDT) from primary swab and urine specimens. In this stud...

Full description

Saved in:
Bibliographic Details
Main Authors: Josephine Moore, Trinity Krueger, Amanda Zapp, Stephen C. Lavey, Kimber L. Munson, Irene A. Stafford, Michael E. Newcomb, Brian Mustanski, Erik Munson
Format: Article
Language:English
Published: American Society for Microbiology 2025-07-01
Series:Microbiology Spectrum
Subjects:
Online Access:https://journals.asm.org/doi/10.1128/spectrum.00738-25
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849429161032024064
author Josephine Moore
Trinity Krueger
Amanda Zapp
Stephen C. Lavey
Kimber L. Munson
Irene A. Stafford
Michael E. Newcomb
Brian Mustanski
Erik Munson
author_facet Josephine Moore
Trinity Krueger
Amanda Zapp
Stephen C. Lavey
Kimber L. Munson
Irene A. Stafford
Michael E. Newcomb
Brian Mustanski
Erik Munson
author_sort Josephine Moore
collection DOAJ
description ABSTRACT Recent work has optimized parameters of a real-time reverse transcriptase PCR-based laboratory-developed test on the Panther Fusion system which detects Mycoplasma genitalium-specific macrolide resistance-associated mutations LDT (MRM-LDT) from primary swab and urine specimens. In this study, MRM-LDT was applied to a large multi-demographic study set to further characterize M. genitalium resistance in the United States. A total of 2,145 primary clinical specimens testing positive for M. genitalium 16S rRNA by transcription-mediated amplification (TMA) were initially titered by the same assay using serial 10-fold dilutions to determine relative target nucleic acid burden. Specimens were then processed for MRM-LDT. Findings were stratified by men who have sex with men (MSM; n = 3 settings), community care (n = 2), and university student (n = 3) populations. The mean log10 target nucleic acid titer of a TMA-positive specimen was 3.46 (median 3; range 0–10). A 43.2% MRM-LDT detection rate was found in specimens derived from community care settings. Analogous assessments of university student and MSM demographics revealed 60.8% and 62.0% detection, respectively (P ≤ 0.0004 versus community care). Within the university student demographic, differences existed in target nucleic acid titer for two settings (P = 0.01); similar differences were encountered between a local MSM cohort and two that were nationally based (P ≤ 0.01). Within the university cohort, MRM-LDT detection rate was increased in symptomatic patients (P = 0.005). M. genitalium macrolide resistance rates among multiple demographics, as determined by MRM-LDT, are high in the United States and are consistent with target nucleic acid burden within the primary specimen. However, caveats experienced within subgroupings of these demographics support reflex MRM-LDT on M. genitalium-positive specimens.IMPORTANCEData generated from a high-throughput, automated system and presented in this report expand upon knowledge of Mycoplasma genitalium-specific macrolide resistance in the United States and may further inform providers on population- or demographic-based considerations for macrolide resistance mutation determination in M. genitalium.
format Article
id doaj-art-a3ba4a90485641a2b4f1bc2ba48d5e41
institution Kabale University
issn 2165-0497
language English
publishDate 2025-07-01
publisher American Society for Microbiology
record_format Article
series Microbiology Spectrum
spelling doaj-art-a3ba4a90485641a2b4f1bc2ba48d5e412025-08-20T03:28:26ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-07-0113710.1128/spectrum.00738-25Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerationsJosephine Moore0Trinity Krueger1Amanda Zapp2Stephen C. Lavey3Kimber L. Munson4Irene A. Stafford5Michael E. Newcomb6Brian Mustanski7Erik Munson8Department of Medical Laboratory Science, Marquette University, Milwaukee, Wisconsin, USADepartment of Medical Laboratory Science, Marquette University, Milwaukee, Wisconsin, USADepartment of Medical Laboratory Science, Marquette University, Milwaukee, Wisconsin, USALoyola University Parkinson School of Health Sciences and Public Health, Maywood, Illinois, USADepartment of Medical Laboratory Science, Marquette University, Milwaukee, Wisconsin, USADepartment of Obstetrics and Gynecology, McGovern Medical School at UTHealth, Houston, Texas, USADivision of Infectious Diseases, Northwestern University, Chicago, Illinois, USADivision of Infectious Diseases, Northwestern University, Chicago, Illinois, USAWisconsin Clinical Laboratory Network Laboratory Technical Advisory Group, Madison, Wisconsin, USAABSTRACT Recent work has optimized parameters of a real-time reverse transcriptase PCR-based laboratory-developed test on the Panther Fusion system which detects Mycoplasma genitalium-specific macrolide resistance-associated mutations LDT (MRM-LDT) from primary swab and urine specimens. In this study, MRM-LDT was applied to a large multi-demographic study set to further characterize M. genitalium resistance in the United States. A total of 2,145 primary clinical specimens testing positive for M. genitalium 16S rRNA by transcription-mediated amplification (TMA) were initially titered by the same assay using serial 10-fold dilutions to determine relative target nucleic acid burden. Specimens were then processed for MRM-LDT. Findings were stratified by men who have sex with men (MSM; n = 3 settings), community care (n = 2), and university student (n = 3) populations. The mean log10 target nucleic acid titer of a TMA-positive specimen was 3.46 (median 3; range 0–10). A 43.2% MRM-LDT detection rate was found in specimens derived from community care settings. Analogous assessments of university student and MSM demographics revealed 60.8% and 62.0% detection, respectively (P ≤ 0.0004 versus community care). Within the university student demographic, differences existed in target nucleic acid titer for two settings (P = 0.01); similar differences were encountered between a local MSM cohort and two that were nationally based (P ≤ 0.01). Within the university cohort, MRM-LDT detection rate was increased in symptomatic patients (P = 0.005). M. genitalium macrolide resistance rates among multiple demographics, as determined by MRM-LDT, are high in the United States and are consistent with target nucleic acid burden within the primary specimen. However, caveats experienced within subgroupings of these demographics support reflex MRM-LDT on M. genitalium-positive specimens.IMPORTANCEData generated from a high-throughput, automated system and presented in this report expand upon knowledge of Mycoplasma genitalium-specific macrolide resistance in the United States and may further inform providers on population- or demographic-based considerations for macrolide resistance mutation determination in M. genitalium.https://journals.asm.org/doi/10.1128/spectrum.00738-25Mycoplasmoides genitaliumtranscription-mediated amplificationmacrolide resistancesemi-quantitation
spellingShingle Josephine Moore
Trinity Krueger
Amanda Zapp
Stephen C. Lavey
Kimber L. Munson
Irene A. Stafford
Michael E. Newcomb
Brian Mustanski
Erik Munson
Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerations
Microbiology Spectrum
Mycoplasmoides genitalium
transcription-mediated amplification
macrolide resistance
semi-quantitation
title Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerations
title_full Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerations
title_fullStr Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerations
title_full_unstemmed Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerations
title_short Automated Mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi-quantitation: patient demographic considerations
title_sort automated mycoplasma genitalium molecular macrolide resistance detection and nucleic acid target semi quantitation patient demographic considerations
topic Mycoplasmoides genitalium
transcription-mediated amplification
macrolide resistance
semi-quantitation
url https://journals.asm.org/doi/10.1128/spectrum.00738-25
work_keys_str_mv AT josephinemoore automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT trinitykrueger automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT amandazapp automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT stephenclavey automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT kimberlmunson automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT ireneastafford automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT michaelenewcomb automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT brianmustanski automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations
AT erikmunson automatedmycoplasmagenitaliummolecularmacrolideresistancedetectionandnucleicacidtargetsemiquantitationpatientdemographicconsiderations