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Detection of genetic mutations associated with macrolide resistance of Mycoplasma pneumoniae

Korean Journal of Pediatrics 2010;53(2):178-183.
Published online February 15, 2010.
Detection of genetic mutations associated with macrolide resistance of Mycoplasma pneumoniae
Chi Eun Oh, Eun Hwa Choi, Hoan Jong Lee
Department of Pediatrics, Seoul National University College of Medicine, Seoul, Korea
Mycoplasma pneumoniae의 macrolide 내성과 연관된 유전자 변이의 검출
오지은, 최은화, 이환종
서울대학교 의과대학 소아과학교실
Correspondence: 
Hoan Jong Lee, Tel: +82.31-787-7283, Fax: +82.31-717-7283, Email: eunchoi@snu.ac.kr
Abstract
Purpose
: The aim of this study was to identify mutations associated with macrolide resistance in Mycoplasma pneumoniae (MP) and to establish a cultural method to determine antimicrobial susceptibility.
Methods
: Nasopharyngeal aspirates (NPAs) were collected from 62 children diagnosed with MP pneumonia by a serologic method or polymerase chain reaction. The 23S rRNA and L4 ribosomal protein genes of MP were amplified and sequenced. To identify mutations in these 2 genes, their nucleotide sequences were compared to those of the reference strain M129. MP cultivation was carried out for 32 (28 frozen and 5 refrigerated) NPAs and M129 strain using Chanock’s glucose broth and agar plate in a 5% CO2 incubator at 37℃ and examined at 2-3 day intervals for 6 weeks.
Results
: Among the 62 specimens, 17 had M144V mutations in ribosomal protein L4. The A2064G mutation was observed in 1 specimen; its 23S rRNA gene was successfully sequenced. Culture for MP was successful from the M129 strain and 2 of the 5 NPAs that were refrigerated for no longer than 3 days. However, MP did not grow from the 28 NPAs that were kept frozen at -80℃ since 2003.
Conclusion
: We found the M144V mutation of L4 protein to be common and that of domain V of 23S rRNA gene was relatively rare among MP. Studies on the prevalence of macrolide-resistant MP and the relationship between the mutations of 23S rRNA gene and ribosomal protein L4 will aid in understanding the mechanism of macrolide resistance in MP.
Key Words: Mycoplasma pneumoniae, Macrolides, Resistance, Mutation


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