title: Evaluation of a real-time fluorescent PCR assay for rapid detection of Group B Streptococci in neonatal blood.
abstract: 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 approximately 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.
authors.affiliation: Pediatric Flight, David Grant USAF Medical Center, Travis Air Force Base, CA 94535, USA. stephen.golden@travis.af.mil
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Golden
authors.forename: David M
authors.initials: DM
authors.lastname: Stamilio
authors.forename: Brian M
authors.initials: BM
authors.lastname: Faux
authors.forename: Wilfred P
authors.initials: WP
authors.lastname: dela Cruz
authors.forename: Craig T
authors.initials: CT
authors.lastname: Shoemaker
authors.forename: Camille L
authors.initials: CL
authors.lastname: Blackmon
authors.forename: Sarah D
authors.initials: SD
authors.lastname: Stassen
authors.forename: Velvet M
authors.initials: VM
authors.lastname: Clark
authors.forename: James W
authors.initials: JW
authors.lastname: Smith
authors.forename: Oswald L
authors.initials: OL
authors.lastname: Johnson
mesh_terms: D001483:Base Sequence; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013290:Streptococcal Infections / Q000097:blood* / Q000175:diagnosis; D013292:Streptococcus agalactiae / Q000302:isolation & purification*; D013997:Time Factors
chemical_list: D004269:DNA, Bacterial