23S核糖体RNA
肺炎支原体
生物
多重聚合酶链反应
单核苷酸多态性
聚合酶链反应
分子生物学
PCR变异
微生物学
基因
基因型
遗传学
核糖核酸
医学
肺炎
核糖体
内科学
作者
Misuk Ji,Nam-Sihk Lee,Jimin Oh,Ji Yoon Jo,Eun Hwa Choi,Soo Jin Yoo,Hyo‐Bin Kim,Sang‐Hyun Hwang,Sang‐Ho Choi,Sang‐Oh Lee,Mi‐Na Kim,Heungsup Sung
标识
DOI:10.1016/j.mimet.2014.04.009
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 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.
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