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, 54 (10), 2563-7

A Novel SimpleProbe PCR Assay for Detection of Mutations in the 23S rRNA Gene Associated With Macrolide Resistance in Mycoplasma Genitalium in Clinical Samples

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A Novel SimpleProbe PCR Assay for Detection of Mutations in the 23S rRNA Gene Associated With Macrolide Resistance in Mycoplasma Genitalium in Clinical Samples

Marianne Gossé et al. J Clin Microbiol.

Abstract

Macrolide-resistant strains of Mycoplasma genitalium are an increasing problem throughout the world, and the implementation of a rapid and sensitive assay for mutation detection to guide treatment is needed. Macrolide-resistant strains have been shown to contain base substitutions in positions 2058 and 2059 (Escherichia coli numbering) in region V of the 23S rRNA gene. In this study, we present a SimpleProbe PCR followed by melting curve analysis to differentiate between macrolide-resistant mutants and wild types. The assay was performed on 159 Mycoplasma genitalium-positive samples, and the results were compared with DNA sequencing. We also looked at the prevalence of macrolide-resistant strains in a Norwegian population. Of 139 samples characterized successfully by sequencing, 54 (39%) were wild types and 85 (61%) were mutants, consisting of 59 (42%) A2059G, 24 (17%) A2058G, 1 (1%) A2058T, and 1 (1%) A2059C mutation. The melting curve analysis correctly differentiated between wild-type and mutant strains in all cases, but it could not identify the different mutant types. The SimpleProbe PCR proved to be a simple, rapid, and reliable method for the detection of macrolide-resistant isolates of Mycoplasma genitalium in a clinical setting.

Figures

FIG 1
FIG 1
Example of melting temperatures of a wild-type strain (64.5°C) and strains with A2058G (58°C) and A2059G (57°C) mutations. The clear distinction of melting temperatures between wild-type and mutant strains is shown.

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This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.

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