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Increased macrolide resistance of Mycoplasma pneumoniae in France directly detected in clinical specimens by real-time PCR and melting curve analysis.
J Antimicrob Chemother. 2009 Jul; 64(1):52-8.JA

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.

Authors+Show Affiliations

Laboratoire de Bactériologie EA 3671, Université Victor Segalen Bordeaux 2 and CHU de Bordeaux, Bordeaux cedex, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Validation Study

Language

eng

PubMed ID

19429926

Citation

Peuchant, O, et al. "Increased Macrolide Resistance of Mycoplasma Pneumoniae in France Directly Detected in Clinical Specimens By Real-time PCR and Melting Curve Analysis." The Journal of Antimicrobial Chemotherapy, vol. 64, no. 1, 2009, pp. 52-8.
Peuchant O, Ménard A, Renaudin H, et al. Increased macrolide resistance of Mycoplasma pneumoniae in France directly detected in clinical specimens by real-time PCR and melting curve analysis. J Antimicrob Chemother. 2009;64(1):52-8.
Peuchant, O., Ménard, A., Renaudin, H., Morozumi, M., Ubukata, K., Bébéar, C. M., & Pereyre, S. (2009). Increased macrolide resistance of Mycoplasma pneumoniae in France directly detected in clinical specimens by real-time PCR and melting curve analysis. The Journal of Antimicrobial Chemotherapy, 64(1), 52-8. https://doi.org/10.1093/jac/dkp160
Peuchant O, et al. Increased Macrolide Resistance of Mycoplasma Pneumoniae in France Directly Detected in Clinical Specimens By Real-time PCR and Melting Curve Analysis. J Antimicrob Chemother. 2009;64(1):52-8. PubMed PMID: 19429926.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Increased macrolide resistance of Mycoplasma pneumoniae in France directly detected in clinical specimens by real-time PCR and melting curve analysis. AU - Peuchant,O, AU - Ménard,A, AU - Renaudin,H, AU - Morozumi,M, AU - Ubukata,K, AU - Bébéar,C M, AU - Pereyre,S, Y1 - 2009/05/09/ PY - 2009/5/12/entrez PY - 2009/5/12/pubmed PY - 2009/7/28/medline SP - 52 EP - 8 JF - The Journal of antimicrobial chemotherapy JO - J Antimicrob Chemother VL - 64 IS - 1 N2 - 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. SN - 1460-2091 UR - https://www.unboundmedicine.com/medline/citation/19429926/Increased_macrolide_resistance_of_Mycoplasma_pneumoniae_in_France_directly_detected_in_clinical_specimens_by_real_time_PCR_and_melting_curve_analysis_ DB - PRIME DP - Unbound Medicine ER -