A Novel SimpleProbe PCR Assay for Detection of Mutations in the 23S rRNA Gene Associated with Macrolide Resistance in Mycoplasma genitalium in Clinical Samples
Autor: | Svein Arne Nordbø, Brita Pukstad, Marianne Gossé, Hilde Lysvand |
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Rok vydání: | 2016 |
Předmět: |
Adult
Male 0301 basic medicine Microbiology (medical) Adolescent Genotyping Techniques 030106 microbiology Population Mutant Mycoplasma genitalium Microbial Sensitivity Tests medicine.disease_cause Polymerase Chain Reaction Melting curve analysis law.invention Young Adult 03 medical and health sciences 23S ribosomal RNA law Drug Resistance Bacterial medicine Humans Transition Temperature Mycoplasma Infections education Polymerase chain reaction education.field_of_study biology Bacteriology Mycoplasma Middle Aged Ribosomal RNA biology.organism_classification Molecular biology Anti-Bacterial Agents RNA Ribosomal 23S Mutation Female Macrolides |
Zdroj: | Journal of Clinical Microbiology |
ISSN: | 1098-660X 0095-1137 |
Popis: | 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. |
Databáze: | OpenAIRE |
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