Real-time PCR assays for the detection and quantification of Streptococcus pneumoniae
Real-time PCR assays for the detection and quantification of Streptococcus pneumoniae
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DOI：
10.1111/j.1574-6968.2010.02044.x
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发表时间：
2010-09-01
影响因子：
2.1
通讯作者：
Kim, Wonyong
中科院分区：
文献类型：
--
作者：
Park, Hee Kuk;Lee, Hee Jung;Kim, Wonyong
肺炎链球菌是全球肺炎的主要病原体。由于草绿色链球菌的成员共享高度的DNA序列同源性，S。来自草绿色组的肺炎是困难的。开发了一种针对芙膜多糖生物合成基因（cpsA）的定量实时PCR检测方法，作为S.肺炎。从135株口腔球菌中提取基因组DNA，进行特异性评价。27个S pneumoniae菌株检测为阳性，而108种其他菌株，包括假链球菌、缓症链球菌和口腔链球菌，未显示出特异性信号。标准曲线的线性回归表明，S. pneumoniae细胞与C（T）值的相关性（R2 = 0.99）。最低检测限为32 fg纯化的基因组DNA，相当于14个S.肺炎。这种新的实时荧光PCR方法可能是非常有用的，作为一种快速和特异性的工具，检测和定量S。肺炎。
Streptococcus pneumoniae is the main etiologic agent of pneumonia worldwide. Because the members of the viridans group streptococci share a high degree of DNA sequence homologies, phenotypic and genotypic discriminations of S. pneumoniae from the viridans group are difficult. A quantitative real-time PCR assay targeting the capsular polysaccharide biosynthesis gene (cpsA) was developed as a species-specific detection tool for S. pneumoniae. The specificity was evaluated using genomic DNAs extracted from 135 oral cocci strains. Twenty-seven S. pneumoniae strains tested positive, whereas 108 other strains including Streptococcus pseudopneumoniae, Streptococcus mitis, and Streptococcus oralis did not show a specific signal. The linear regression of standard curves indicated high correlations between the log numbers of S. pneumoniae cells and the C(T) values (R2=0.99). The minimal limit of detection was 32 fg of purified genomic DNA, equivalent to 14 genomes of S. pneumoniae. This new real-time PCR method may be very useful as a rapid and specific tool for detecting and quantifying S. pneumoniae.