Single-nucleotide polymorphism PCR for the detection of Mycoplasma pneumoniae and determination of macrolide resistance in respiratory samples

被引：17

作者：

Ji, Misuk ^{[1
,2
]}

Lee, Nam-Sihk ^{[3
]}

Oh, Ji-Min ^{[3
]}

Jo, Ji Yoon ^{[3
]}

Choi, Eun Hwa ^{[4
]}

Yoo, Soo Jin ^{[5
]}

Kim, Hyo-Bin ^{[6
]}

Hwang, Sang-Hyun ^{[7
,8
]}

Choi, Sang-Ho ^{[2
,9
]}

Lee, Sang-Oh ^{[2
,9
]}

Kim, Mi-Na ^{[1
,2
]}

Sung, Heungsup ^{[1
,2
]}

机构：

[1] Asan Med Ctr, Dept Lab Med, Seoul, South Korea

[2] Univ Ulsan, Coll Med, Seoul, South Korea

[3] SolGent Co Ltd, Taejon, South Korea

[4] Seoul Natl Univ Hosp, Dept Pediat, Seoul 110744, South Korea

[5] Inje Univ, Coll Med, Sanggye Paik Hosp, Dept Lab Med, Seoul, South Korea

[6] Inje Univ, Coll Med, Sanggye Paik Hosp, Dept Pediat, Seoul, South Korea

[7] Natl Canc Ctr, Res Inst & Hosp, Dept Lab Med, Goyang Si, South Korea

[8] Natl Canc Ctr, Res Inst & Hosp, Hematol Malignancy Branch, Goyang Si, South Korea

[9] Asan Med Ctr, Dept Infect Dis, Seoul, South Korea

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2014年 / 102卷

关键词：

Mycoplasma pneumoniae; Macrolide resistance; Single-nucleotide polymorphism PCR; 23S rRNA gene; Mutation; COMMUNITY-ACQUIRED PNEUMONIA; REAL-TIME PCR; MOLECULAR ANALYSIS; PEDIATRIC-PATIENTS; CHINA; SUSCEPTIBILITY; INFECTION; SPECIMENS; SHANGHAI; CHILDREN;

D O I：

10.1016/j.mimet.2014.04.009

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 73 M. pneumoniae-positive samples and 100 M. 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. (C) 2014 Elsevier B.V. All rights reserved.

引用

页码：32 / 36

页数：5

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