Article

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.

Download full-text

Full-text

Available from: Cécile Bébéar, Aug 21, 2015
0 Followers
 · 
324 Views
  • Source
    • "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). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The aim of this study was to develop a single-nucleotide polymorphism (SNP) PCR assay to be performed directly on respiratory samples for the simultaneous detection of Mycoplasma pneumoniae and its 23S rRNA gene mutations, which are responsible for macrolide resistance. For multiplex SNP PCR, two outer primers for amplification of the 23S rRNA gene and two mutant-specific primers for the discrimination of single base changes were designed. A total of 73M. pneumoniae-positive samples and 100M. pneumoniae-negative samples were analyzed using this assay. By SNP PCR, we detected two mutations conferring high-level macrolide resistance in 22 samples (A2063G from 20 and A2064G from 2 samples); these results are identical to those produced by the 23S rRNA gene sequencing of M. pneumoniae-positive samples. Thus, this assay can be used as a practical method for the simultaneous detection of M. pneumoniae and mutations associated with macrolide resistance directly from respiratory samples.
    Journal of microbiological methods 04/2014; 102. DOI:10.1016/j.mimet.2014.04.009 · 2.10 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Not Available
    18th Annual Symposium on Frequency Control. 1964; 02/1964
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: A two-dimensional, steady-state thermal conduction analysis is made on a finite, plane homogeneous medium (a printed circuit board, or PCB) to examine the trace behavior. The trace is modeled as a zero-thickness, strip heat source with specified uniform temperature, and its position in the medium is varied. A two-dimensional thermal analysis is also performed on a multilayered cell model with finite heat source to establish an accurate, effective thermal conductivity for a typical PCB
    Semiconductor Thermal Measurement and Management Symposium, 1991. SEMI-THERM VII. Proceedings., Seventh Annual IEEE; 03/1991
Show more