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

Laboratoire de Bactériologie EA 3671, Université Victor Segalen Bordeaux 2 and CHU de Bordeaux, Bordeaux cedex, France.
Journal of Antimicrobial Chemotherapy (Impact Factor: 5.44). 06/2009; 64(1):52-8. DOI: 10.1093/jac/dkp160
Source: PubMed

ABSTRACT 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.
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.
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.
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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Available from: Cécile Bébéar, Aug 21, 2015
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    • "Previous studies have confirmed that a single base mutation at position 2063 in domain V of the 23S rRNA gene of M. pneumoniae is the most prevalent mutation, followed by a mutation at position 2064, both conferring high-level macrolide resistance. Current molecular techniques for identifying these mutations include direct sequencing, restriction fragment length polymorphism analysis, real-time PCR with high-resolution melting curve analysis, PCR with pyrosequencing, and allele-specific PCR (Bebear et al., 2011; Li et al., 2012; Matsuoka et al., 2004; Peuchant et al., 2009; Spuesens et al., 2010, 2012; Wolff et al., 2008). These methods all have various advantages and limitations (Li et al., 2012). "
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