Real-time PCR assays for the detection and quantification of Streptococcus pneumoniae

FEMS Microbiology Letters
H K ParkW Kim

Abstract

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 (R(2)=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.

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Citations

Jan 29, 2013·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·S EspositoN Principi
Mar 20, 2012·The Pediatric Infectious Disease Journal·Susanna EspositoUNKNOWN Italian Pneumococcal CAP Group
Feb 7, 2012·Applied and Environmental Microbiology·Hee Kuk ParkWonyong Kim
Oct 9, 2012·Journal of Microbiology, Immunology, and Infection = Wei Mian Yu Gan Ran Za Zhi·Chen-Yin LaiLuan-Yin Chang
Dec 18, 2015·Future Microbiology·Rachid Y YahiaouiEllen E Stobberingh
Jul 6, 2013·Scandinavian Journal of Infectious Diseases·Vesna Cvitkovic SpikManica Müller Premru
May 28, 2016·Archives of Oral Biology·Cyntia Ferreira RibeiroAna Christina Claro Neves
May 27, 2017·Analytical Chemistry·Jinping WangDavid Tai Leong
Jun 19, 2018·Emerging Infectious Diseases·Catarina Silva-CostaUNKNOWN Portuguese Study Group of Invasive Pneumococcal Disease of the Pediatric Infectious Disease Society
Mar 19, 2019·Indian Journal of Medical Microbiology·Sreeram Chandra Murthy PeelaB Adhishivam
May 22, 2021·Transactions of the Royal Society of Tropical Medicine and Hygiene·Seok Hwee KooThean Yen Tan

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