Increased macrolide resistance of Mycoplasma pneumoniae in France directly detected in clinical specimens by real-time PCR and melting curve analysis

O. Peuchant, A. Ménard, H. Renaudin, Miyuki Morozumi, K. Ubukata, C. M. Bébéar, Sabine Pereyre

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

Objectives: Mycoplasma pneumoniae is a common aetiological agent of community-acquired respiratory tract infections for which macrolides are the treatment of choice. In France, only two macrolide-resistant isolates were reported in 1999. In contrast, several recent data reported that macrolide-resistant M. pneumoniae isolates have been spreading since 2000 in Japan. Mutations A2058G (Escherichia coli numbering), A2058C, A2059G, A2062G, C2611A and C2611G in domain V of the 23S rRNA gene were associated in vivo or in vitro with this resistance. The aim of this study was to determine whether macrolide resistance of M. pneumoniae is emerging in France. Patients and methods: We developed a duplex real-time PCR for the detection of the six 23S rRNA mutations associated with macrolide resistance in M. pneumoniae and a simplex real-time PCR for the identification of the A2058G mutation, the most common one. Both methods rely on fluorescence resonance energy transfer coupled to melting curve analysis and are directly applicable to clinical samples. The duplex real-time PCR assay, first validated on 40 genetically characterized M. pneumoniae strains, was then applied directly on 248 French respiratory tract clinical samples. Results: Among M. pneumoniae-positive specimens collected before 2005, no macrolide-resistant M. pneumoniae isolate was detected. In contrast, among 51 samples collected between 2005 and 2007, five (9.8%) yielded a resistant genotype, suggesting a recent increase in macrolide-resistant M. pneumoniae isolates in France. Conclusions: The epidemiological monitoring of macrolide resistance in this species has become necessary in France and Europe, and will be made easier by using these PCR assays.

Original languageEnglish
Pages (from-to)52-58
Number of pages7
JournalJournal of Antimicrobial Chemotherapy
Volume64
Issue number1
DOIs
Publication statusPublished - 2009
Externally publishedYes

Fingerprint

Mycoplasma pneumoniae
Macrolides
Freezing
France
Real-Time Polymerase Chain Reaction
Mutation
Epidemiological Monitoring
Community-Acquired Infections
Fluorescence Resonance Energy Transfer
rRNA Genes
Respiratory Tract Infections
Respiratory System
Japan
Genotype
Escherichia coli
Polymerase Chain Reaction

Keywords

  • 23S rRNA
  • Antimicrobial resistance epidemiology
  • Laboratory methods
  • Low respiratory tract infections
  • LRT
  • Target gene mutation

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Increased macrolide resistance of Mycoplasma pneumoniae in France directly detected in clinical specimens by real-time PCR and melting curve analysis. / Peuchant, O.; Ménard, A.; Renaudin, H.; Morozumi, Miyuki; Ubukata, K.; Bébéar, C. M.; Pereyre, Sabine.

In: Journal of Antimicrobial Chemotherapy, Vol. 64, No. 1, 2009, p. 52-58.

Research output: Contribution to journalArticle

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abstract = "Objectives: Mycoplasma pneumoniae is a common aetiological agent of community-acquired respiratory tract infections for which macrolides are the treatment of choice. In France, only two macrolide-resistant isolates were reported in 1999. In contrast, several recent data reported that macrolide-resistant M. pneumoniae isolates have been spreading since 2000 in Japan. Mutations A2058G (Escherichia coli numbering), A2058C, A2059G, A2062G, C2611A and C2611G in domain V of the 23S rRNA gene were associated in vivo or in vitro with this resistance. The aim of this study was to determine whether macrolide resistance of M. pneumoniae is emerging in France. Patients and methods: We developed a duplex real-time PCR for the detection of the six 23S rRNA mutations associated with macrolide resistance in M. pneumoniae and a simplex real-time PCR for the identification of the A2058G mutation, the most common one. Both methods rely on fluorescence resonance energy transfer coupled to melting curve analysis and are directly applicable to clinical samples. The duplex real-time PCR assay, first validated on 40 genetically characterized M. pneumoniae strains, was then applied directly on 248 French respiratory tract clinical samples. Results: Among M. pneumoniae-positive specimens collected before 2005, no macrolide-resistant M. pneumoniae isolate was detected. In contrast, among 51 samples collected between 2005 and 2007, five (9.8{\%}) yielded a resistant genotype, suggesting a recent increase in macrolide-resistant M. pneumoniae isolates in France. Conclusions: The epidemiological monitoring of macrolide resistance in this species has become necessary in France and Europe, and will be made easier by using these PCR assays.",
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