This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wolff, B. J.
Right arrow Articles by Winchell, J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wolff, B. J.
Right arrow Articles by Winchell, J. M.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, October 2008, p. 3542-3549, Vol. 52, No. 10
0066-4804/08/$08.00+0     doi:10.1128/AAC.00582-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Detection of Macrolide Resistance in Mycoplasma pneumoniae by Real-Time PCR and High-Resolution Melt Analysis{triangledown}

Bernard J. Wolff, W. Lanier Thacker, Stephanie B. Schwartz, and Jonas M. Winchell*

Respiratory Disease Branch, Centers for Disease Control and Prevention, Atlanta, Georgia

Received 2 May 2008/ Returned for modification 24 June 2008/ Accepted 11 July 2008

Mycoplasma pneumoniae is a significant cause of community-acquired pneumonia, which is often empirically treated with macrolides or azalides such as erythromycin or azithromycin. Recent studies have discovered the existence of macrolide-resistant strains within the population that have been mapped to mutations within the domain V region of the 23S rRNA gene. Currently, identification of these resistant strains relies on time-consuming and labor-intensive procedures such as restriction fragment length polymorphism, MIC studies, and sequence analysis. The current study reports two distinct real-time PCR assays that can detect the A2063G or A2064G base mutation (A2058G or A2059G by Escherichia coli numbering) conferring macrolide resistance. By subjecting the amplicon of the targeted domain V region of the 23S rRNA gene to a high-resolution melt curve analysis, macrolide-resistant strains can quickly be separated from susceptible strains. Utilizing this method, we screened 100 clinical isolates and found 5 strains to possess mutations conferring resistance. These findings were concordant with both sequencing and MIC data. This procedure was also used successfully to identify both susceptible and resistant genotypes in 23 patient specimens. These patient specimens tested positive for the presence of M. pneumoniae by a separate real-time PCR assay, although the bacteria could not be isolated by culture. This is the first report of a real-time PCR assay capable of detecting the dominant mutations that confer macrolide resistance on M. pneumoniae, and these assays may have utility in detecting resistant strains of other infectious agents. These assays may also allow for clinicians to select appropriate treatment options more rapidly and may provide a convenient method to conduct surveillance for genetic mutations conferring antibiotic resistance.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, 1600 Clifton Rd. N.E., MS: G-03, Atlanta, GA 30333. Phone: (404) 639-4921. Fax: (404) 639-4215. E-mail: jwinchell{at}cdc.gov

{triangledown} Published ahead of print on 21 July 2008.

Antimicrobial Agents and Chemotherapy, October 2008, p. 3542-3549, Vol. 52, No. 10
0066-4804/08/$08.00+0     doi:10.1128/AAC.00582-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Peuchant, O., Menard, A., Renaudin, H., Morozumi, M., Ubukata, K., Bebear, 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. J Antimicrob Chemother 64: 52-58 [Abstract] [Full Text]  
  • Waites, K. B., Crabb, D. M., Duffy, L. B. (2009). Comparative In Vitro Susceptibilities of Human Mycoplasmas and Ureaplasmas to a New Investigational Ketolide, CEM-101. Antimicrob. Agents Chemother. 53: 2139-2141 [Abstract] [Full Text]  
  • Liu, Y., Ye, X., Zhang, H., Xu, X., Li, W., Zhu, D., Wang, M. (2009). Antimicrobial Susceptibility of Mycoplasma pneumoniae Isolates and Molecular Analysis of Macrolide-Resistant Strains from Shanghai, China. Antimicrob. Agents Chemother. 53: 2160-2162 [Abstract] [Full Text]  
  • Xin, D., Mi, Z., Han, X., Qin, L., Li, J., Wei, T., Chen, X., Ma, S., Hou, A., Li, G., Shi, D. (2009). Molecular Mechanisms of Macrolide Resistance in Clinical Isolates of Mycoplasma pneumoniae from China. Antimicrob. Agents Chemother. 53: 2158-2159 [Abstract] [Full Text]  
  • Mitchell, S. L., Wolff, B. J., Thacker, W. L., Ciembor, P. G., Gregory, C. R., Everett, K. D. E., Ritchie, B. W., Winchell, J. M. (2009). Genotyping of Chlamydophila psittaci by Real-Time PCR and High-Resolution Melt Analysis. J. Clin. Microbiol. 47: 175-181 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»