Rapid detection of Streptococcus pneumoniae by real-time fluorescence quantitation PCR

doi:10.3969/j.issn.1673-8640.2013.03.014

Abstract

Abstract: Objective　To establish real-time fluorescence quantitation polymerase chain reaction (PCR) for the detection and epidemiological studies of Streptococcus pneumoniae. Methods　The autolysin (lyt) gene and hemolysin (ply) gene sequences were selected to develop primers and probe. A total of 10 strains of Streptococcus pneumoniae，13 strains of non-Streptococcus pneumoniae DNA and the different concentration strains of Streptococcus pneumoniae DNA were detected by real-time fluorescence quantitation PCR，and 200 clinical specimens were detected by primers and probe. The specificity and sensitivity were also analyzed. Results　The 10 strains of Streptococcus pneumoniae were measured to obtain amplification products. However，13 strains of non-Streptococcus pneumoniae DNA had no obvious amplification，and the sensitivity was 100fg. Among the 200 clinical specimens，real-time fluorescence quantitation PCR detected 42 cases of Streptococcus pneumoniae-positive (the positive rate was 21%)，while the culture method detected 16 cases of Streptococcus pneumoniae-positive (the positive rate was 8%). Conclusions　Real-time fluorescence quantitation PCR is a rapid，sensitive and specific assay for the detection of Streptococcus pneumoniae. It can be used for the diagnosis and epidemiological studies.

Key words: Streptococcus pneumoniae, Real-time fluorescence quantitation polymerase chain reaction, Taqman probe

YAN Shanhuo，SUN Lei，FU Kepeng，ZHUO Yongguang，YANG Shanye，FAN Zuqian，HUANG Yongxia. Rapid detection of Streptococcus pneumoniae by real-time fluorescence quantitation PCR[J]. , 2013, 28(3): 221-224.

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