Evaluation of a real-time fluorescent PCR assay for rapid detection of Group B Streptococci in neonatal blood

Streptococcus agalactiae (Group B Streptococcus: GBS) is the major causative agent of neonatal sepsis. Neonates at risk for GBS infections are empirically administered broad-spectrum antibiotics for at least 48 h pending blood culture results. A rapid assay to expedite detection of GBS would facilit...

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Bibliographic Details
Published in:Diagnostic microbiology and infectious disease 2004-09, Vol.50 (1), p.7-13
Main Authors: Golden, Stephen M., Stamilio, David M., Faux, Brian M., dela Cruz, Wilfred P., Shoemaker, Craig T., Blackmon, Camille L., Stassen, Sarah D., Clark, Velvet M., Smith, James W., Johnson, Oswald L.
Format: Article
Language:English
Subjects:
Bacteriology
Base Sequence
Biological and medical sciences
Blotting, Southern
DNA, Bacterial - analysis
Electrophoresis, Agar Gel
Female
Fundamental and applied biological sciences. Psychology
Humans
Infant, Newborn
Infectious diseases
Male
Medical sciences
Microbiology
Miscellaneous
Molecular Sequence Data
Reverse Transcriptase Polymerase Chain Reaction
Sensitivity and Specificity
Streptococcal Infections - blood
Streptococcal Infections - diagnosis
Streptococcus agalactiae
Streptococcus agalactiae - isolation & purification
Time Factors
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Summary:Streptococcus agalactiae (Group B Streptococcus: GBS) is the major causative agent of neonatal sepsis. Neonates at risk for GBS infections are empirically administered broad-spectrum antibiotics for at least 48 h pending blood culture results. A rapid assay to expedite detection of GBS would facilitate initiation of specific antibiotic therapy. Conversely, expeditious proof of absence of infection will avoid unnecessary antibiotic use. Using the LightCycler™, we evaluated a hybridization probe polymerase chain reaction (PCR) assay to detect GBS-specific cfb gene target DNA sequence in blood specimens. Both sensitivity and specificity of the real-time PCR assay was 100%. The assay demonstrated 100% specificity when tested against 26 non-GBS bacteria. This method is capable of detecting as few as ∼100 copies or 10 pg of GBS genomic DNA. This real-time PCR method is rapid, sensitive, and specific for the detection of GBS in neonatal blood samples and holds great promise in its utility in the diagnostic laboratory.
ISSN:0732-8893
1879-0070
DOI:10.1016/j.diagmicrobio.2004.04.021